转蜘蛛Spidroin1基因绵羊早期克隆胚的研究
|
摘要
蜘蛛丝无与伦比的生物学特性决定了其在应用领域具有广阔的应用前景。以生物学方法获取具有蜘蛛丝特性的活性蛋白,亦或是通过转基因的方法直接获得类似蜘蛛丝的动物纤维,一直以来是许多科学研究者的奋斗目标。本研究利用蜘蛛拖丝蛋白基因四聚体4S、pcDNA3.1和pIRES2-EGFP载体,在构建pcDNA3.1-4S和pIRES2-EGFP-4S表达载体的基础上,以脂质体转染法将蜘蛛拖丝蛋白(Spidroin1)基因转染细胞并获得转基因阳性细胞株；以转基因阳性细胞为核供体,采用体细胞核移植的方法获得转蜘蛛Spidroinl基因绵羊克隆胚；采用PCR方法检测转基因克隆胚的蜘蛛拖丝蛋白(Spidroinl)基因整合状况。本研究的结果如下：
1. pcDNA3.1-4S及pIRES2-EGFP-4S表达载体的构建利用pcDNA3.1载体和蜘蛛拖丝蛋白基因四聚体4S构建了pcDNA3.1-4S表达载体,BgtlⅡ, BamHI双酶切及PCR鉴定显示,蜘蛛拖丝蛋白基因4S已连接到pcDNA3.1-4S中；利用pIRES2-EGFP载体和蜘蛛拖丝蛋白基因4S构建了pIRES2-EGFP-4S, XhoⅠ、BglⅡ双酶切及PCR鉴定显示,蜘蛛拖丝蛋白基因4S连接到pIRES2-EGFP中。
2.转蜘蛛Spidroin1基因绵羊成纤维细胞系的建立将0.44g/mL经线性化的pcDNA3.1-4S和pIRES2-EGFP-4S,通过脂质体法分别转染利用常规方法经原代和传代培养的绵羊成纤维细胞,用含有抗生素和含400μg/mL G418的培养基进行筛选,获得转pcDNA3.1-4S和pIRES2-EGFP-4S的阳性细胞；对获得的转基因阳性细胞进行形态学观察、生长曲线、倍增时间等生物学特性检测结果,符合正常成纤维细胞的特性。
3.绵羊卵母细胞获取方法的筛选通过比较抽吸、切割、抽吸结合切割的方法对绵羊的获卵效果,其结果,与抽吸和切割法相比,划线法辅助的切割法,能够获得最多的A、B级卵母细胞(约8个/每个卵巢)。
4.绵羊卵母细胞体外成熟将A、B级卵母细胞以TCM199-HCO3+2.2g/mL NaHCO3+5μg/mL FSH+1IU/mL LH+1μg/mL雌二醇+50μg/mL庆大霉素+10%FBS+0.38mmol/L丙酮酸钠+10mmol/L Hepes为培养液,对绵羊卵母细胞进行体外成熟培养,可以获得83.4%的最高成熟率。表明该成熟液适合用于绵羊卵母细胞的体外成熟。
5.成熟卵母细胞孤雌激活体外成熟绵羊卵母细胞用5μmol IA23187激活5min后,再用2mmol/L的6-DMAP培养4h,能够获得最高91.5%的激活率,激活后的孤雌胚经培养后84.7%胚胎发育至桑椹胚。本研究结果显示,IA23187联合6-DMAP的激活方法可用于绵羊体外成熟卵母细胞的孤雌激活。
6.转基因阳性细胞的体细胞核移植利用上述研究中建立的体细胞核移植程序,分别以绵羊成纤维细胞和转蜘蛛拖丝蛋白基因细胞进行体细胞核移植,其结果,正常绵羊体细胞融合率(72.7%),与其相比,转基因细胞的融合率(70.7%),两者差异不显著,表明外源基因的转入对早期核移植重构胚的发育能力没有显著影响。
7.转蜘蛛拖丝蛋白基因阳性细胞的体细胞重构胚的发育能力
将采用体细胞核移植方法获得的转蜘蛛拖丝蛋白基因重构胚进行体外培养,其结果与正常的绵羊成纤维细胞通过体外发育桑椹胚的比率(13.8%)相比,转基因阳性细胞的体细胞的核移植的体外发育至桑椹胚的比率(11.6%),两者之间的差异不显著；将获得的转蜘蛛拖丝蛋白基因融合重构胚直接移入未妊娠的受体绵羊输卵管中,观察到受体妊娠,但未能获得转蜘蛛拖丝蛋白基因的绵羊后代。
综上所述,本研究利用所构建的蜘蛛拖丝蛋白基因表达载体pcDNA3.1-4S和pIRES2-EGFP-4S,获得转基因阳性细胞；利用转基因阳性细胞,采用体细胞核移植方法建立了获取转蜘蛛拖丝蛋白基因重构胚的方法；获得的重构胚具有体外发育至桑椹胚的能力。本研究为进一步开展被毛中表达蜘蛛拖丝蛋白的转基因绵羊相关研究奠定了坚实的基础。
The unapproachable biological characteristics of spider silk determines it has broad application in many fields. To obtain the active protein with spider silk characteristics using biological method, or animal fibers like spider silk using transgenic approach,is some scientific researchers'goal all these years.This study utilized the spider dragline silk protein gene4S、 pcDNA3.1and pIRES2-EGFP vector to contruct pcDNA3.1-4S and pIRES2-EGFP-4S expression vector,utilized the liposomal and linearized spider dragline silk protein gene4S to transfect sheep fibroblasts,then obtained the positive cell line;used the positive cell line as donor nuclear to obtain trans-Spidroinl gene sheep reconstruceted embryos by nuclear transfer; identified whether the the spider dragline silk protein gene integrated into the reconstruceted embryos.The results of this study were showed as follows.
1. Construction of pcDNA3.1-4S and pIRES2-EGFP-4S expression vector
The pcDNA3.1-4S expression vector was constructed by pcDNA3.1vector and spider dragline silk protein gene4S, the result showed that the spider dragline silk protein gene4S had integrated into the pcDNA3.1-4S expression vector after BglⅡ、BamHI double enzymes cutted and PCR identificated;the pIRES2-EGFP-4S vector was constructed by spider dragline silk protein gene4S and pIRES2-EGFP vector, the result showed that the spider dragline silk protein gene4S had integrated into the pIRES2-EGFP-4S vector after XhoⅠ, BglⅡ double enzymes cutted and PCR identificated.
2. Establishment of trans-Spidroninl gene sheep fibroblasts cell line
Utilized the liposomal to transfect the sheep primary and passage fibroblast with0.4μg/mL linearized pcDNA3.1-4S and pIRES2-EGFP-4S,screened the sheep fibroblasts by culture media with400μg/mL G418and antibiotics, and finally obtained trans-pcDNA3.1-4S and pIRES2-EGFP-4S cell line; observed the morphology,drawed the growth curve,calculated the proliferation time of the transgene cell,all the results were accord to the normal fibroblast feature.
3. Screened the method of sheep oocytes collecting
Compared the effect of sheep oocytes collecting method by suction,cut and suction combined with cut, the results showed that the cut assisted with lineation can obtain about8oocytes (including A and B levels) each ovary.
4. Vitro maturation of sheep oocytes
Used the media:TCM199-HCO3+2.2g/mL NaHCO3+5μg/mL FSH+1IU/mL LH+1μg/mL E2+50μg/mL Gentamicin+10%FBS+0.38mmol/L sodium pyruvate+10mmol/L Hepes as culture media to culture A and B levels of sheep oocytes,obtained the highest83.4%maturation rate,it showed that this culture media were fit for sheep oocytes vitro maturation.
5. Parthenogenetic activation of sheep matured oocytes
The vitro matured sheep oocytes were firstly activated with5μmol IA23187,then cultured in2mmol/L6-DMAP for4h,it could obtain91.5%activation rate,after cultured84.7%of the parthenogenetic activation embryos were developed into morula. it showed that IA23187combined with6-DMAP were fit for parthenogenetic activation of matured sheep oocytes.
6. Somatic cells nuclear transfer of transgene cell
Used the sheep fibroblasts and trans-spider dragline silk protein gene cell as the donor nuclear,after somatic cells nuclear transfer operation which were founded before,the results showed that the abnormal cell melt rate was72.7%,the melt rate of transgene cell was70.7%,there were no difference,it indicated that there were no influence of the exogenous gene integrated into the reconstruceted embryos on it's development ability.
7. Development ability of trans-spider dragline silk protein gene reconstruceted embryos by nuclear transfer
Cultured the trans-spider dragline silk protein gene reconstruceted embryos by nuclear transfer,the morula rate of transgene cell as donor nuclear was11.6%,moreover the morula of abnormal cell as donor nuclear wasl3.8%,there were no difference between them transferred the trans-spider dragline silk protein gene reconstruceted embryos into the oviduct of the donor sheep,pregnance was observered,but no trans-spider dragline silk protein gene lamb obtained.
In summary,this study used the pcDNA3.1-4S and pIRES2-EGFP-4S expression vector were constructed to obtain the transgene cell line;used the transgene cell as donor nuclear to obtain trans-spider dragline silk protein gene reconstruceted embryos, the reconstruceted embryos obtained had the ability developing into morula.this study built the foundation for further research of obtaining trans-spider dragline silk protein gene sheep in hair.
1. pcDNA3.1-4S及pIRES2-EGFP-4S表达载体的构建利用pcDNA3.1载体和蜘蛛拖丝蛋白基因四聚体4S构建了pcDNA3.1-4S表达载体,BgtlⅡ, BamHI双酶切及PCR鉴定显示,蜘蛛拖丝蛋白基因4S已连接到pcDNA3.1-4S中；利用pIRES2-EGFP载体和蜘蛛拖丝蛋白基因4S构建了pIRES2-EGFP-4S, XhoⅠ、BglⅡ双酶切及PCR鉴定显示,蜘蛛拖丝蛋白基因4S连接到pIRES2-EGFP中。
2.转蜘蛛Spidroin1基因绵羊成纤维细胞系的建立将0.44g/mL经线性化的pcDNA3.1-4S和pIRES2-EGFP-4S,通过脂质体法分别转染利用常规方法经原代和传代培养的绵羊成纤维细胞,用含有抗生素和含400μg/mL G418的培养基进行筛选,获得转pcDNA3.1-4S和pIRES2-EGFP-4S的阳性细胞；对获得的转基因阳性细胞进行形态学观察、生长曲线、倍增时间等生物学特性检测结果,符合正常成纤维细胞的特性。
3.绵羊卵母细胞获取方法的筛选通过比较抽吸、切割、抽吸结合切割的方法对绵羊的获卵效果,其结果,与抽吸和切割法相比,划线法辅助的切割法,能够获得最多的A、B级卵母细胞(约8个/每个卵巢)。
4.绵羊卵母细胞体外成熟将A、B级卵母细胞以TCM199-HCO3+2.2g/mL NaHCO3+5μg/mL FSH+1IU/mL LH+1μg/mL雌二醇+50μg/mL庆大霉素+10%FBS+0.38mmol/L丙酮酸钠+10mmol/L Hepes为培养液,对绵羊卵母细胞进行体外成熟培养,可以获得83.4%的最高成熟率。表明该成熟液适合用于绵羊卵母细胞的体外成熟。
5.成熟卵母细胞孤雌激活体外成熟绵羊卵母细胞用5μmol IA23187激活5min后,再用2mmol/L的6-DMAP培养4h,能够获得最高91.5%的激活率,激活后的孤雌胚经培养后84.7%胚胎发育至桑椹胚。本研究结果显示,IA23187联合6-DMAP的激活方法可用于绵羊体外成熟卵母细胞的孤雌激活。
6.转基因阳性细胞的体细胞核移植利用上述研究中建立的体细胞核移植程序,分别以绵羊成纤维细胞和转蜘蛛拖丝蛋白基因细胞进行体细胞核移植,其结果,正常绵羊体细胞融合率(72.7%),与其相比,转基因细胞的融合率(70.7%),两者差异不显著,表明外源基因的转入对早期核移植重构胚的发育能力没有显著影响。
7.转蜘蛛拖丝蛋白基因阳性细胞的体细胞重构胚的发育能力
将采用体细胞核移植方法获得的转蜘蛛拖丝蛋白基因重构胚进行体外培养,其结果与正常的绵羊成纤维细胞通过体外发育桑椹胚的比率(13.8%)相比,转基因阳性细胞的体细胞的核移植的体外发育至桑椹胚的比率(11.6%),两者之间的差异不显著；将获得的转蜘蛛拖丝蛋白基因融合重构胚直接移入未妊娠的受体绵羊输卵管中,观察到受体妊娠,但未能获得转蜘蛛拖丝蛋白基因的绵羊后代。
综上所述,本研究利用所构建的蜘蛛拖丝蛋白基因表达载体pcDNA3.1-4S和pIRES2-EGFP-4S,获得转基因阳性细胞；利用转基因阳性细胞,采用体细胞核移植方法建立了获取转蜘蛛拖丝蛋白基因重构胚的方法；获得的重构胚具有体外发育至桑椹胚的能力。本研究为进一步开展被毛中表达蜘蛛拖丝蛋白的转基因绵羊相关研究奠定了坚实的基础。
The unapproachable biological characteristics of spider silk determines it has broad application in many fields. To obtain the active protein with spider silk characteristics using biological method, or animal fibers like spider silk using transgenic approach,is some scientific researchers'goal all these years.This study utilized the spider dragline silk protein gene4S、 pcDNA3.1and pIRES2-EGFP vector to contruct pcDNA3.1-4S and pIRES2-EGFP-4S expression vector,utilized the liposomal and linearized spider dragline silk protein gene4S to transfect sheep fibroblasts,then obtained the positive cell line;used the positive cell line as donor nuclear to obtain trans-Spidroinl gene sheep reconstruceted embryos by nuclear transfer; identified whether the the spider dragline silk protein gene integrated into the reconstruceted embryos.The results of this study were showed as follows.
1. Construction of pcDNA3.1-4S and pIRES2-EGFP-4S expression vector
The pcDNA3.1-4S expression vector was constructed by pcDNA3.1vector and spider dragline silk protein gene4S, the result showed that the spider dragline silk protein gene4S had integrated into the pcDNA3.1-4S expression vector after BglⅡ、BamHI double enzymes cutted and PCR identificated;the pIRES2-EGFP-4S vector was constructed by spider dragline silk protein gene4S and pIRES2-EGFP vector, the result showed that the spider dragline silk protein gene4S had integrated into the pIRES2-EGFP-4S vector after XhoⅠ, BglⅡ double enzymes cutted and PCR identificated.
2. Establishment of trans-Spidroninl gene sheep fibroblasts cell line
Utilized the liposomal to transfect the sheep primary and passage fibroblast with0.4μg/mL linearized pcDNA3.1-4S and pIRES2-EGFP-4S,screened the sheep fibroblasts by culture media with400μg/mL G418and antibiotics, and finally obtained trans-pcDNA3.1-4S and pIRES2-EGFP-4S cell line; observed the morphology,drawed the growth curve,calculated the proliferation time of the transgene cell,all the results were accord to the normal fibroblast feature.
3. Screened the method of sheep oocytes collecting
Compared the effect of sheep oocytes collecting method by suction,cut and suction combined with cut, the results showed that the cut assisted with lineation can obtain about8oocytes (including A and B levels) each ovary.
4. Vitro maturation of sheep oocytes
Used the media:TCM199-HCO3+2.2g/mL NaHCO3+5μg/mL FSH+1IU/mL LH+1μg/mL E2+50μg/mL Gentamicin+10%FBS+0.38mmol/L sodium pyruvate+10mmol/L Hepes as culture media to culture A and B levels of sheep oocytes,obtained the highest83.4%maturation rate,it showed that this culture media were fit for sheep oocytes vitro maturation.
5. Parthenogenetic activation of sheep matured oocytes
The vitro matured sheep oocytes were firstly activated with5μmol IA23187,then cultured in2mmol/L6-DMAP for4h,it could obtain91.5%activation rate,after cultured84.7%of the parthenogenetic activation embryos were developed into morula. it showed that IA23187combined with6-DMAP were fit for parthenogenetic activation of matured sheep oocytes.
6. Somatic cells nuclear transfer of transgene cell
Used the sheep fibroblasts and trans-spider dragline silk protein gene cell as the donor nuclear,after somatic cells nuclear transfer operation which were founded before,the results showed that the abnormal cell melt rate was72.7%,the melt rate of transgene cell was70.7%,there were no difference,it indicated that there were no influence of the exogenous gene integrated into the reconstruceted embryos on it's development ability.
7. Development ability of trans-spider dragline silk protein gene reconstruceted embryos by nuclear transfer
Cultured the trans-spider dragline silk protein gene reconstruceted embryos by nuclear transfer,the morula rate of transgene cell as donor nuclear was11.6%,moreover the morula of abnormal cell as donor nuclear wasl3.8%,there were no difference between them transferred the trans-spider dragline silk protein gene reconstruceted embryos into the oviduct of the donor sheep,pregnance was observered,but no trans-spider dragline silk protein gene lamb obtained.
In summary,this study used the pcDNA3.1-4S and pIRES2-EGFP-4S expression vector were constructed to obtain the transgene cell line;used the transgene cell as donor nuclear to obtain trans-spider dragline silk protein gene reconstruceted embryos, the reconstruceted embryos obtained had the ability developing into morula.this study built the foundation for further research of obtaining trans-spider dragline silk protein gene sheep in hair.
引文
[1]邵敬党。蜘蛛丝的性能特征分析。棉纺织技术,2005,33(11)：653～657
[2]Vollrath F. Strength and structure of spider'silks. Biological Macromolecules,2000,74:67-83
[3]何兰芝,陈莉萍,王学梅。二十一世纪新型纤维——蜘蛛丝。甘肃科技,2004,20(11)：133～134
[4]S.F.Y.Li et al.Comparative Study of the Internal Structure of Kelvar and Spider Silk by Atomic Force Microscopy.J.Vac.Sci.Technol.A.,1994(4):1891-1894
[5]S.F.Y.Li et al.New Internal Structure of Spider Silk Revealed by Atomic Force Microscopy. Biophysical Journal,1994 (66):1209-1212
[6]Kitagawa et al.Microstructure of Spider Dragline.J.Soc.Mat.Sci.,2001(50):1213-1217
[7]Augsten K et al. Glycoproteins and Skin-core Structure in Nephlia Clavipes Spider Silk Observed by Light and Electron.Scanning,2000(1):12-15
[8]S.Frische et al.Elongate Cavities and Skin-core Structure in Nephlia Spider Silk Observed by Electron Microscopy. J. Microscopy,1998(189):64-74
[9]Z.Shao et al.Heterogeneous Morphology of Nephila Edulis Spider Silk and its Significance for Mechanical Properties. Polymer,1999 (40):4709-4711
[10]Sirichaisit J, Yong R J, Vollrath F. Molecular deformation in spider dragline silk subjected to stress. Polymers,1999,40:2493-2500
[11]Riekel C, Muller M, Vollrath F. In-situ X-ray diggraction studies during forced silking of a Nephila senigalensis spider. Macromocules,1999,32:4464-4466
[12]Hijirida H, Do K G, Michal, et al. C-13 NMR of Nephial clavipes major ampulllate silk gland. Biophys,1996,71:3442-3447
[13]Shao Z Z, Vollrath F, Sirichaisit J. Analysis of spider silk in native and supercontracted states using Raman spectroscopy. Polymer,1999,40:2493-2500
[14]刘海洋,工伟霞,刘长军等。纺织新材料——蜘蛛丝。纺织导报,2004(1)：28—30
[15]周文聪,张贤,张敏等。影响蜘蛛丝性能的因素及开发技术。纺织科技进展,2010(4)：58-61
[16]赵博。新一代高性能纤维—蜘蛛丝纤维。国外丝绸,2005,(1)：22-23
[17]刘庆生,段亚峰。蜘蛛丝的结构性能与研究现状。四川丝绸,2005,2：16-18
[18]袁辉。蜘蛛丝的研究与开发利用。国外纺织技术,2006,6：7-9
[19]蒋平,肖永红,吕太勇等。蜘蛛丝的结构与机械性能研究进展。井冈山大学学报,2010,31(4)：37-43
[20]陈瑶,孟清,卿凤翎。蜘蛛丝纤维的特性与开发生产。现代纺织技术,2006,6：53-56
[21]J. Gatesyetal. Extreme diversity conservation and convergence of spider silk fibroin sequences. Science,2001,291:2603-2605
[22]段亚峰,冀勇斌。蜘蛛丝开发应用的现状与进展。丝绸,2002,7：46-47
[23]王琳纲,胡平。蜘蛛丝的研究进展及展望。新材料产业,2007,9：63～67
[24]J. Gatesyetal. Extreme diversity, Conservation, and convergence of spider silk fibroin sequences. Science,2001,291:2603-2605
[25]F. Vollrath and D. P. Knight. Liquid Crystalline Soinning of Spidersilk. Nature,2001,410: 545-548
[26]黄献聪,施楣梧。蜘蛛丝的力学性能及其应用取向。纺织导报,2004,3：32-38
[27]潘志娟,邱芯薇。蜘蛛丝的物理性能研究。苏州大学学报,2003：23(1)：18-22
[28]刘辉芬,蒋平,肖永红。两种不同功能蜘蛛丝的物理化学结构表征与力学性。武汉大学学报,2010,56(1)：93-99
[29]盛家墉,潘志娟,陈宇岳。蜘蛛丝的化学组成与结构初探,2000,4：8-13
[30]刘全勇,江雷。仿生学与天然蜘蛛丝仿生材料。高等学校化学学报,2010,31(6)：1065-1071
[31]张慧勤,王志新。蜘蛛丝的研究与应用。中原工学院学报,2005,16(4)：47-50
[32]Xu M, Lewis R V. Structure of a protein superfiber:spider dragline silk. Proc Natl Acad Sci U S A,1990,87(18):7120-7124
[33]Hinman M B, Lewis R V. Isolation of a clone encoding a second dragline silk fibroin. J Biol Chem,1992,267 (27):19320-19324
[34]Colgin M A, Lewis R V. Spider minor ampullate silk proteins contain new repetitive sequences and highly conserved non-silk-like "spacer regions". Protein Sci,1998, 7(3):667-672
[35]Beckwitt R,Arcidiacono S.Sequecn conservation in the C-teminal region of spider silk proteins(Spidroin) from Nehpila clavipes(Tetragnathidae) and Araneus bicentenarius(Araneidae).J.Biol.Chem; 1994,269:6661-6663
[36]Hayashi C Y, Lewis R V. Evidence from flagelliform silk cDNA for the structural basis of elasticity and modular nature of spider silk. J Mol Biol,1998,275(5):773-784
[37]Guerette P,Ginzinger D,Weber B,Gosline J. Silk properties determined by gland-specific expression of a spider fibroin gene family.Science,1996.272:112-115
[38]Prince J T, Mcgrath K P, DiGirolamo C M, et al. Construction, cloning and expression of synthetic genes encoding spider dragline silk. Biochemistry,1995, 34(34):10879-10885
[39]Xia X X, Qian Z G, Ki C S, et al. Native-sized recombinant spider silk protein produced in metabolically engineered Escherichia coli results in a strong fiber. Proc Natl Acad Sci U S A,2010,107(32):14059-14063
[40]Fahnestock R, Bedzyk L A. Production of synthetic spider dragline silk protein in picha pastoris. Appl Microbiol Biothechnol,1997,47(1):33-39
[41]Scheller J, Guhrs K H, Grosse F, et al. Production of spider silk proteins in tobacco and potato. Nat Biotechnol,2001,19(6):573-577
[42]Scheller J, Henggeler D, Viviani A, et al. Purification of spider silk elastin from transgenic plants and application for human chondrocyte proliferation. Transgenic Res, 2004,13(1):51-57
[43]Piruzian E S, Bogush V G, Sidoruk K V, et al. Construction of synthetic genes for analogs of spider silk spidroin 1 and their expression in tobacco plants. Mol Biol, 2003,37(4):554-560
[44]Barr L A, Fahnestock S R, Yang JJ, et al. Production and purification of recombinant DP1B silk-like protein in plants. Mol Breeding,2004,13(4):345-356
[45]Lazaris A, Arcidiacono S, Huang Y, et al. Spider silk fiber spun from soluble recombinant silk produced in mammalian cells. Science,2002,295(5554):472-476
[46]Gatesy J, Hayashi C, Motriuk D, et al. Extreme diversity, conservation, and convergence of spider silk fibroin sequences. Science,2001,291(5513):2603-260519324
[47]Xu H T, Fan B L, Yu S Y, et al. Construct synthetic gene encoding artificial spider dragline silk protein and its expression in milk of transgenic mice. Anim Biotechnol, 2007,18(1):1-12
[48]Ponder BA. Cancer genetics. Nature.2001.411(6835):336-341
[49]梁洋。蜘蛛拖丝蛋白基因Spidroinl嵌合体小鼠的研究。东北林业大学博士论文.2011,6
[50]Carl A, Pinkert. Transgenic Animal Technology:A Laboratory Handbook. Second Edition. Chenical Industry Press,2004,134-135
[51]Buchschacher Jr GL.Introduction to retroviruses and retroviral vectors. Somat Cell Mol Genet,2001,26:1-11
[52]Hu W S and Pathak V K. Design of Retroviral Vectors and Helper Cells for Gene Therapy. Pharmacol Rev Dec,2000,52(4):493-511
[53]唐东红,代解杰,仝品芬等。泡沫病毒载体的研究进展。病毒学报,2000,9(3)：282-284
[54]Yu S F, Sullivan M D, Linial M L. Evidence that the human foamy virus genome is DNA. J Virol,1999,73:1565-1572
[55]许建,李世崇,陈昭烈。逆转录病毒表达系统及其在外源蛋白高效表达中的应用。 中国生物工程杂志,2008,28(5)：116-121
[56]王振发,王烈,卫立辛。基因治疗病毒载体的研究进展,2007,4(3)：490-492
[57]Dalba C, Bellier B, Kasahara N et al..Replication-competent Vectors and Empty Virus-like Particles:New Retroviral Vector Designs for Cancer Gene Therapy or Vaccines.The American Society of Gene. Therapy Mol Ther,2007,15(3):457-466
[58]Yu S S, Han E, Iiong Y et al. Construction of a retroviral vector production system with the minimum possibility of a homologous recombination. Gene Ther,2003,10:706-711
[59]Dillon P, Lenz J, Rosen C. Construction of a replication-competent murine retrovirus vector expressing the human immunodeficiency virus type 1 tat transactivator protein. J Virol,1991,65:4490-4493
[60]Solly S, Trajcevski S, Frisen C et al. Replicative retroviral vectors for cancer gene therapy. Cancer. Gene Ther,2003,10:30-39
[61]Wang W, Tai C, Kasahara N et al. Highly efficient and tumor-restricted gene transfer to malignant gliomas by replication-competent retroviral vectors. Hum Gene Ther,2003,14:117-127
[62]Trajcevski S, Solly S, Frisen C et al. Characterization of a semi-replicative gene delivery system allowing propagation of complementary defective retroviral vectors. J Gene Med,2005,7:276-287
[63]Doan L, Li M, Chen C et al. Virus-like particles as HIV-1 vaccines. Rev Med Virol,2005,15:75-78
[64]Robertson E, Bradley A, Kuehn M, et al. Germ-line transmission of genes introduced into cultured pluripotential cells by retroviral vector. Nature,1986,323(6087):445-448
[65]Chan A W. Homan E J, Ballou L U et al. Transgenic cattle produced by reverse-transcribed gene transfer in oocytes. Proc Natl Acad Sci USA,1998,95(24):14028-14033
[66]Chan A W, Chong K Y, Martinovich C et al. Transgenic Monkeys Producedby Retroviral Gene Transferinto Mature Oocytes. Science,2001,291(5502):309-312
[67]Matsunar H, Onodera M, Tada N et al. Transgenic-Cloned Pigs Systemically Expressing Red Fluorescent Protein, Kusabira-Orange. Cloning Stem Cells,2008,10(3):313-323
[68]Benigni A and Remuzzi G. Gene Therapy for Renal Diseases and Transplantation. Contrib Nephrol Basel Karger,2008,159:30-46
[69]Rainov N G, Kramm C M, Banning U et al. Immune response induced by retrovirus-mediated HSV-tk/GCV pharmacogene therapy in patients with glioblastoma multiforme. Gene Ther,2000,7(21):1853-1858
[70]谢文练,黄健,韩金利等。逆转录病毒介导HSV-TK基因结合更昔洛韦对裸鼠膀胱癌模型的抑癌作用。中华泌尿外科杂志,2005,26(11)：752-754
[71]Hauster H,Gross G.Brunsw, et al. Inducibility of humanB-Interferon gene in mouse L-cell clones. Nature,1982,297:650
[72]孔永,秦秀云。动物细胞培养技术研究进展。重庆文理学院学报,2007,26(4)：54-56
[73]谢文忠。巯嘌吟键连卟啉抗癌药物的合成、表征及生物活性研究。湖南大学硕士学位论文,2008,19
[74]肖深初。三嗪卟啉衍生物的合成、表征及其生物活性研究。湖南大学博士学位论文,2009,63
[75]魏宗波。雏鸡心肌细胞原代培养及锰对肉仔鸡心肌细胞MnSOD基因表达调控研究。西北农林科技大学,2008,3
[76]冯为慧。淡水鱼类细胞现采现培技术的初步建立与关键技术点研究。南昌大学硕士学位论文,2012,10
[77]顾涵英,冯佑民。无血清细胞培养基及其主要补充因子。生物化学与生物物理进展,1990,17(4)：255-259
[78]马忠仁,冯玉萍,李倬。动物血清在细胞培养中的重要性及其质量控制标准。西北民族大学学报(自然科学版)。2003,24(50)：56-58
[79]刘艳。细胞培养及常见问题的处理。山东畜牧兽医,2008,29：13-16
[80]甘露,张志辉,吕美娜。细胞培养技术。中国组织工程研究与临床康复,2008,12(29)：5739-5742
[81]王立新,杨朝霞。动物细胞培养及应用。黄牛杂志,2000,26(3)：45-48
[82]桂东城,蒋涛。转基因克隆动物的研究进展。中国畜牧兽医,2009,36(11)：114-117
[83]骆继怀,独军政,常惠芸。转基因克隆动物的发展及其应用。动物医学进展,2010,31(7)：103-106
[84]刘西梅,乔宪凤,张立苹等。转基因动物研究的技术思路。湖北农业科学,2012,51(21)：4837-4840
[85]Gordon J W,Scangos G A,Plotrin D J,et al.Genetic transformation of mouse embryo by microinjection of purified DNA.Proc Natl A-cad Sci USA,1980,77:7380-7384
[86]余华,叶健强,刘丹。动物转基因及克隆技术研究。信阳农业高等专科学校学报,2009,19(2)：104-108
[87]刘晓兰,王仁华,工峥等。动物转基因技术及其应用。湖南饲料,2010,1：28-31
[88]Evans M J,Kaufman M H. Establishment in culture of pluripotential cells from mouse embryos. Nature,1981,292:154-156
[89]刘黎青,李娟,周盛年。胚胎干细胞及在生殖医学中的应用。中国组织工程研究与临床康复,2011,15(1)：113-115
[90]张瑞杰,苗向阳。动物转基因技术研究现状与应用前景。中国细胞生物学报,2010,32(5)：800-807
[91]王玉琴。绵羊同期发情研究进展。畜禽业,2003,4：4-6
[92]蒋长炜,冯国献。绵羊同期发情技术概述。畜牧与饲料科学。2010,31(10)：109-110
[93]李武,王丽娟,李丽华。绵羊胚胎移植研究进展。黑龙江畜牧兽医,2002,7：48-50
[94]傅文栋,李风德,牛建克。绵羊胚胎移植研究进展。黑龙江动物繁殖。2010,18(5)：20-21
[95]Gordon JB, Uehlinger V.Fertile intestine nuclei.Nature.1966,210:1240-1241
[96]McGrath J and Sloter D.Inability of mouse blastomere nuclei transtected to enucleated zygotes to support development in vitro.Science,1984,226:1317-1319
[97]朵红。动物核移植技术研究概况。2005,35(4)：43～44
[98]孙志宏,邱瑞,李延清等。哺乳动物核移植研究。安徽农业科学,2008,36(7)：2783-2784,2788
[99]Wilmut I, Schnieke A E, McWhir J, et al. Viable offspring derived from fetal and adult mammalian cells. Nature,1997,385:810-813
[100]Baguisi A, Behboodi E, Melican D T, et al. Production of goats by somatic cell nuclear transfer. Nat Biotechnol,1999,17 (5):456-461
[101]Kubota C, Yamakuchi H, Todoroki J, et al. Six cloned calves produced from adult fibroblast cells after long-term culture. Proc Natl Acad Sci,2000,97(3):990-995
[102]Dai Y, Vaught T D, Boone J, et al. Targeted disruption of the alphal,3-galactosyl transferase gene in cloned pigs Nat Biotecnol,2002,20(3):251-255
[103]Wakayama T, Rodriguez I, Perry A C, et al. Mice cloned from embryonic stem cells. Proc Natl Acad Sci USA..1999,96(26):14984-14989
[104]Chesne P, Adenot P G, Viglietta C, et al. Cloned rabbits produced by nuclear transfer from adult somatic cells. Nat Biotechnol,2002,20(4):366-369
[105]Shin T, Kraemer D, Pry or J, et al. A cat cloned by nuclear transplantation. Nature, 2002,415 (6874):859
[106]Lagutina I, Lazzari G, Duchi R, et al. Somatic cell nuclear transfer in horses:effect of oocyte morphology, embryo reconstruction method and donor cell type. Reproduction,2005,130(4):559-567
[107]杨彩侠,陈大元。动物克隆机理研究进展。动物学研究,2003,24(1)：73-78
[108]Kubota C, Yamakuchi H, Todoroki J, et al. Six cloned calves produced from adult fibroblast cells after long-term culture.Proc Nat Acad Sci USA,2000,97(3):990-995
[109]Enright B P, Jeong B S, Yanget X, et al. Epigenetic characteristics of bovine donor cells for nuclear transfer:levelsof histone acetylation.Biology of Reproduction,2003,69(5):1525-1530
[110]Poehland R, Al-Rostum F, Becker F, et al. Donor cell lines considerably affect the outcome of somatic nuclear transfer in the case of bovines. Journal of Reproduction and Development,2007,53 (4):737-748
[111]Betthauser J, Forsberg E, Augenstein M, et al. Production of cloned pigs from in vitro systems. Nat Biotechnol,2000,18(10):1055-1059
[112]Hill J R, Winger Q A, Burghardt R C, et al. Bovine nuclear transfer embryo development using cells derived from a cloned fetus. Anim Reprod Sci,2001,67(1 12):17-26
[113]Takahashi K, Yamanaka S. Induction of pluripotent stem cells frommouse embryonic and adult fibroblast cultures by defined factors. Cell,2006,126(4):663-679
[114]Schwarzer M, Carnwath J W, Lucas-Hahn A, et al. Isolation of bovine cardiomyocytes for reprogramming studies based on nuclear transfer. Cloning Stem Cells,2006,8(3):150-158
[115]Mizutani E, Ono T, Li C, et al. Propagation of senescent mice using nuclear transfer embryonic stem cell lines. Genesis,2008,46(9):478-483
[116]张传山,郭毅,谷瑞环等。成体兔转基因成纤维细胞的克隆分离及其核移植研究。生物化学与生物物理进展,2009(9)：1186-1192
[117]Tao Y, Han W, Zhang M, et al. Production of Boer goat (Capra hircus) by nuclear transfer of cultured and cryopreserved fibroblast cells into slaughterhouse derived oocytes. Czech Journal of Animal Science,2009,54 (10):448-460
[118]工建锋,张勇,赵兴绪。不同采卵方式对绵羊卵母细胞质量、体外成熟及体外受精的影响。动物医学进展,2007,28(8)：51-54
[119]万鹏程,张云生,石文艳。绵羊卵母细胞不同采集方法对体外成熟培养的影响。石河子大学学报,2004,22(5)：401-404
[120]Keskintepel L, Darwish G M, Kenimer A T, et al.Term development of caprine embryos derived from immature oocytes in vitro.Theriogenology,1994,42(3):527-535
[121]许杰,马恒东,吴兴杰。采集方法、EGF对山羊卵母细胞体外成熟的影响。安徽农业科学,2007,35(17)：5185-5186,5191
[122]刘青春,李武峰,岳文斌等。不同卵巢运输温度和培养方法对卵母细胞体外成熟的影响。畜牧兽医杂志,2008,27(4)：16-17
[123]汪立芹,杨梅,陈童等。季节和方法对牛、绵羊卵母细胞回收及体外成熟的影响。中国畜牧兽医,2008,35(11)：74-76
[124]张伟,工利红。影响卵母细胞体外成熟的因素。黄牛杂志,2005,31(1)：51-54
[125]Mayes MA, Sirard MA. The influence of cumulus oocyte complex morphology and meiotic inhibitors on the kinetics of nuclear maturation cattle. Theriogenology, 2000,54:911-922
[126]Masui Y and markert C L.Cytoplasmic centrol of nuclear behavior during meiotic muturation of frog oocytes.J Exp Zool,1971,177:129-146
[127]Smith L D, Ecker.The interaction of steroids with Rena pipiensoocytes in the induction of maturation.Dev Biol,1971,25:232-247
[128]Wakayama T, Perry A C F, Zoccotti M, et al.Full-tem development of mice from enucleated oocytes injected with cumulus cell nuclei.Nature,1998,394 (6691):369-374
[129]Zhou Q, Jouneau A, Brochard V, et al.Development potential of mouse embryos reconstructed from metaphase embryonic stem cell nuclei.Biol Reprod,2001,65 (2):412-419
[130]Lanza R P, Cibelli J B, Diaz C, et al.Cloning of endangered species (Bos Gaurus) using interspecies nuclear transfer.Bio.Reprod.,2001,64:1487-1493
[131]Kono T, Obata Y, Wu Q, et al. Birth of parthenogenetic mice that can develop to adulthood.Nature,2004,428 (6985):860-864
[132]李清。卵母细胞去核方法及其对卵母细胞发育的影响。山东畜牧兽医,2007,28(6)：42-44
[133]Wilmut I, Schnieke A E,McWhir J,et al. Viable offspring derives from fetal and adult mammalian cells.Nature,1997,385:810-813
[134]Illmensee K,Hoppe PC.Nuclear transplantationin mouse musculus.development potential of nuclei from preimplantation embryos.Cell,1981,23:9-16
[135]Willadsen SM. Nuclear transplantation in sheep embryo. Nature,1986,320:63-65
[136]Prather RS, Barnes FL, Sims MM, et al.Nuclear transplantation in the bovine embryos: Assessment of donor nuclei and recipient oocyte. Biol Reprod,1987,37:859-866
[137]Stice SL, Robl JM. Nuclear reprogramming in nuclear transplant rabbit embryo. Biol Repod,1988,39:657-664
[138]Prather RS, Sims MM, First NL. Nuclear transplantation in early pig embryos. Biol Repod,1989,44:414-418
[139]杜淼,陆德裕,沈三兵。家兔早期胚胎细胞发育能力的研究。实验生物学报,1990,23(4)：477-482
[140]张涌,王建辰,钱菊汾等。山羊卵核移植的研究。中国农业科学,1991,24(5)：1-6
[141]邹贤刚,李光三,王玉阁。山羊胚胎细胞经继代细胞核移植后其发育能力的研究。科学通报,1995,40(3)：264-267
[142]李雪峰,石德顺,谭丽玲等。电激活对完全体外化牛细胞核移植的影响。畜牧兽医学报,1996,27：495-500
[143]赵浩斌,陈乃清,魏庆信等。猪卵核移植的研究。武汉大学学报,1997,43 (4)：505-510
[144]Li M, John J, Richard L, et al. Rhesus monkey produced by nuclear transfer [J]. Biol Reprod,1997,57:454-459
[145]Wakayama T,Perry ACF, Zoccotti M et al. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature,1998,394(7):369-374
[146]Cibelli J B,Stice S L,Golueke P J,et al. Cloned transgenic calves produced from nonquiescent fetal fibroblasts.Science,1998,280:1256-1258
[147]132.Vignon X,Chesns P,LebourhisD.Dvelopmental Potential of bovine embryos reconstructed with somatic nuclei from cultured skin and muscle fetal cells.Theriogenology,1998,49:392
[148]KatoY,Tani T,Sotomaru Y,etal.Eight calves cloned from somatic cells of a single adult.Science,1998,282:2095-2098
[149]Well D N,Misica P M,Tervit H R,etal.Production of cloned calves following nuclear transfer with cultured adult mural granulose cells.Biology of ReProduetion,1999,60:996-1005
[150]Baguisi A, Behboodi E, Melican DT, Pollock JS, Destrempes MM, Cammuso C, Williams JL, Nims SD, Porter CA, Midura P, Palacios MJ, Ayres SL, Denniston RS, Hayes ML, Ziomek CA, Meade HM, Godke RA, Gavin WG, Overstrom EW,Echelard Y. Production of goats by somatic cell nuclear transfer.Nat Biotechnol,1999;17(5):456-461
[151]工玉阁,邹贤刚,成国祥等。由胎儿成纤维细胞而来的克隆山羊。科学通报,1999,44(21)：2319-2323
[152]OnishiA,Iwamoto M,Akita J,etal.pigs cloning by microinjeetion of fetal fibroblast nuclei.Science.2000,289:1187-1193
[153]McCreath K J, Howcroft J, Campbell K H,et al.Production of gene-targeted sheep by nucleartransfer from cultured somatic cells.Nature,2000,405:1066-1069
[154]Chesne P, Adenot P G, Viglietta,et al. Cloned rabbits produced by nuclear transfer from adult somatic cells.Nat Biotechnol,2002,20(4):366-369
[155]Li, Z. Sun, X. Chen, J.etal.Cloned ferrets produced by somatic cell nuclear transfer. Dev Biol.2006. (293):2
[156]Nisar A.Wani, U.Wernery,F.A.H.Hassan,4R.et al.Production of the First Cloned Camel by Somatic Cell Nuclear Transfe. Biol Reprod,2010,82:373-379
[157]Briggs R,King T J.Transplantation of living cell nuclei from blastula cells into enucleated frogps eggs.Proc Natl Acad Sci USA,1952,38:455-463
[158]童第周,吴尚勤,叶毓芬等。鱼类细胞核的移植。科学通报,1963,7：60-61
[159]童第周,叶毓芬,陆德裕等。鱼类不同亚科间的细胞核移植。动物学报,1973,19(3)：201-209
[160]McGrath JD,etal.Inability of mouse blastomere nuclei transferred to enucleated zyzotes tosupport development in vitro.Scienee,1984,226:1317
[161]WolfeBA,etal.Methods in bovine nuclear transfer.Theriogenology,1992,37:5
[162]梅祺,邹贤刚,杜森。鼠兔核质杂交胚胎早期发育的研究。实验生物学报,1993：26(4)：389-397
[163]陈大元,孙青原,刘冀珑等。大熊猫供核体细胞在兔卵胞质中可去分化而支持早期重构胚的发育。中国科学(C辑),1999,29(3)：324-330
[164]Dominko T,Mitalipova M,Haley B,et al.Bovine oocytes cytoplasm supports development of embryos produced by nuclear transfer of somatic cell nuclei from various mammalian species.Biology of Reproduction,1999,60:1496-1502
[165]Lanza RP, Cibelli JB, Diaz F, et al, Cloning of an endangered species (Bos gaurus) using interspecies nuclear transfer. Cloning 2000,2(2):79-90
[166]Murakami M, Otoi T, Wongsrikeao P, et al.Development of interspecies cloned embryos in yak and dog.Cloning Stem Cells,2005,7(2):77-81
[167]Thongphakdee A, Numchaisrika P, Omsongkram S, et al.In vitrodevelopment of marbled cat embryos derived from inter-species somatic cell nuclear transfer. Reprod Domest Anim,2006,41 (3):219-226
[168]Sugimura S., Narita K., Yamashiro H., Sugawara A., Shoji T.,Terashita Y., Nishimori K., Konno T., Yoshida M., and Sato E., Interspecies somatic cell nucleus transfer with porcine oocytes as recipients:A novel bioassay system for assessing the competence of canine somatic cells to develop into embryos, Theriogenology,2009,72(4):549-559
[169]Sha H.Y., Chen J.Q., Chen J., Zhang P.Y., Wang P., Chen L.P.,Cheng GX., and Zhu J.H., Fates of donor and recipient mitochondrial DNA during generation of interspecies SCNT-derived human ES-like cells, Cloning and Stem Cells,2009,11(4):497-507
[170]舒金辉,陶锋,张波。体细胞核移植表观遗传的研究进展。中国比较医学杂志,2008,18(8)：49-52
[171]刘秀丽。哺乳动物核移植技术研究进展。贵州畜牧兽医,2006,30(2)：8-11
[172]周艳华,贾银海,苏雷等。影响细胞核移植效率的因素。现代畜牧兽医,2008,2：46-50
[173]王建,向舒,陆凤花等。哺乳动物体细胞核移植后核重编程的研究进展。安徽农业科学,2010,38(25)：13732-13733
[174]汪立芹,王静,赵云程等。供核细胞及处理方式对绵羊核移植效率的影响。华北农学报,2012,27：69-73
[175]王春生。转拟蜘蛛拖丝蛋白基因绵羊成纤维细胞系的建立。东北农业大学博士论文。2007,55
[176]原璐。转拟蜘蛛拖丝蛋白基因四聚体绵羊成纤维细胞株的筛选。东北林业大学硕 士论文。2009,20
[177]薛庆善。体外培养的原理与技术,北京：科学出版社,2001,2-6
[178]Kazutoshi Takahashi, Shinya Yamanaka. Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors. Cell,2006,126:663-676
[179]Haslam S Z. Progesterone effects on deoxyribonucleic acid synthesis in normal mouse mammary glands. Endocrinology.1988,122(2):464-470
[180]李扬,吴凯峰,郭旭东等。脂质体介导外源基因体外转染牛胎儿成纤维细胞条件的优化。遗传,2002,24(6)：653-655
[181]佟慧丽。奶山羊乳腺上皮细胞系的建立。东北农业大学硕士论文,2007,11
[182]刘亚,章孝荣,张运海等。牛、羊成纤维细胞的冷冻与冷藏保存。中国兽医学报,2004,24(2)：197-198
[183]翟秀岩,马文锋,热海佐保子。培养细胞的冷冻保存和解冻复苏的操作方法。中国医科大学学报,2002,31(1)：77-78
[184]王晓玉,柳增善,任洪林。“生物刚性及弹性蛋白”—蜘蛛丝研究进展。第四军医大学学报,23：31-34
[185]Hinman, M B, Jones, J.A.&Lewis R V. Synthetic spider silk:a modular fibre. TIBTECH, 2000,18:374-379
[186]Scheller J, Guhrs KH, Grosse F et al. Production of spider silk proteins in tobacco and potato. Nat Biotechnol,2001,19:573-577
[187]Pelletier J, Flynn M E, Kaplan G, Racaniello V, Sonenberg N. Mutational analysis of upstream AUG codons of poliovirus RNA. J Virol.1988,62(12):4486-4492
[188]郑月茂。转基因山羊乳腺上皮细胞系建立与转基因克隆胚发育。西北农林科技大学博士论文,2005,132
[189]王沛涛,何立群,舒志全等。细胞低温生物学研究进展。齐鲁医学杂志,2005,20：1-3
[190]潘琼,陈彦,梅黎等。冷冻复苏对大鼠干细胞代谢活性的影响。中国药理学与毒理学杂志,2008,22(6)：457-462
[191]杜晨光。绵羊卵母细胞孤雌激活与体细胞核移植的研究。内蒙古大学硕士学位论文,2005,7
[192]孙杰。影响绵羊卵丘细胞核移植效果主要因素的研究。河北农业大学硕士学位论文,2008,17.
[193]赖彦。绵羊核移植技术的研究。内蒙古农业大学硕士学位论文,2010,18
[194]南木甲,张雁平。不同采卵方法及卵巢所处性周期阶段对绵羊卵母细胞体外成熟的影响。中国畜牧兽医,2007,34(7)：137-139
[195]张永胜,工宏伟,皮文辉。两种采卵方法对绵羊卵母细胞体外受精的影响。甘肃 畜牧兽医,2005,3：9-11
[196]Sommersberg B,Bulling A,SalzerU,et al.Gap junction communication and connexin 43 gene expression in a rat granulose cell line:regulation by follicle-stimulating hormone.BioReprod,2000,63(6):1661-1668
[197]周平,刘东军,闫真等。TGFα、EGF对绵羊卵母细胞成熟的影响。细胞生物学杂志,2007,29：713-717
[198]郭延华。EGF对不同直径卵泡绵羊卵母细胞体外成熟、受精及胚胎发育的影响。石河子大学硕士学位论文,2010,2
[199]毛晓霞,万双秀。生殖激素对绵羊卵母细胞体外成熟的影响。上海畜牧兽医通讯。2009,1：33-34
[200]张晓建,安志兴,李学斌。卵母细胞体外成熟时间对绵羊核移植效率的影响。安徽农业科学,2008,36(32)：14120-14121,14132
[201]Albertini D F, Combelles C M, Benecchi E, et al. Cellular basis for paracrine regulation of ovarian follicle development。Reproduction,2001,121,647-653
[202]赛务加甫。提高绵羊体细胞核移植效率的研究。西北农林科技大学博士论文,2007,41
[203]曲杨燕,赵雁伟,李凯等。FSH对绵羊卵母细胞体外核成熟的影响。中国畜牧兽医,2008,35(12)：80-83
[204]郭宪,陈学进,叶荣等。牛卵泡卵母细胞体外成熟的研究。中国草食动物,2004,24(2)：3-6
[205]林雍峰,赵兴绪,张勇。绵羊卵母细胞核成熟、卵丘扩展与激素浓度关系的研究。甘肃农业大学学报,2012,47(1)：22-27
[206]梁素丽,李向臣,白修云等。影响卵母细胞体外成熟的因素。中国畜牧兽医,2007,34(9)：36-39
[207]何良军,彭莉,马兵祥等。绵羊卵母细胞最优运输温度和体外成熟时间的研究。塔里木大学学报,2005,17(4)：5-7
[208]乔利敏,郭彤,乔富强。绵羊卵巢运输、保存温度及时间对其卵母细胞孤雌发育能力的影响。浙江农业学报,2012,24(4)：563—566
[209]王旭光。绵羊卵母细胞体外成熟和孤雌激活的研究。新疆农业大学硕士论文。2003,27
[210]王建辰,章孝荣。动物生殖调控。安徽科学技术出版社,1998：220～230
[211]Machaty Z, PratherR S. Strategies for activating nuclear transfer oocytes. Reprod Fertil Dev,1998,10:599-613
[212]Tateno H, Kamiguchi Y.1997. Parthenogenetic activation of Chinese hamster oocytes by chemical stimuli and its cytogenetic evaluation. Mol Reprod Dev.47:7278
[213]Morgan A and Jacob R. Ionomycin enhances influx by stimulating store-regulated cation entry not by a direct action at the plasma membrane. Bio Chem J.1994,300:665 672
[214]Wang WH, Machaty Z, Ruddoch N, et al.Activation of porcine oocytes with alcium ionophore:effects of extracellularcalcium.Mol Reprod,1999 (1):99-107
[215]施巧婷,王旭光,王立虎。中国畜牧杂志。2004,40(6)：10-13
[216]张晓建。绵羊卵母细胞孤雌激活与体细胞核移植。西北农林科技大学硕士论文,2005,30
[217]彭礼繁,罗光彬。牛、羊卵母细胞孤雌激活的研究。畜禽业,2008,226：56-58
[218]周欢敏,李宝山等。不同激活方法对绵羊体外成熟卵母细胞孤雌生殖的影响。内蒙古农业大学学报,2008,29(1)：78-81
[219]张云生。绵羊卵母细胞孤雌激活及冷冻研究。石河子大学硕士论文。2005,22
[220]陆凤花,舒金辉,黎江等。哺乳动物克隆电融合技术的影响因素。广西农业生物科学,2007,26(2)：179-182
[221]王立虎,黄俊成,杨梅。绵羊体细胞核移植实验研究。草食家畜,2001,111：17-19
[222]工亮,彭涛,朱海等。电融合条件对新疆盘羊(Ovis ammon)和转基因(GFP)萨能奶山羊核移植胚胎发育的影响。西北农业学报,2006,15(2)：1-6
[223]H.R.TERVIT,D.G.WHITTINGHAM and L.E.A.ROWSON.Successful culture in vitro of sheep and cattle ova.J.Reprod.Fert.1972,30,493-497
[224]Grazyna Ptak,Pasqualino Loi,Maria Dattena,et al.Offspring from One-Month-Old Lambs: Studies on the Developmental Capability of Prepubertal Oocytes. Biology of reproduction,1999,61:1568-1574
[225]Pasqualino Loi,Michael Clinton,Barbara Barboni, et al.Nuclei of Nonviable Ovine Somatic Cells Develop into Lambs after Nuclear Transplantation. Biology of reproduction, 2002:67,126-132
[226]GUO Jitong, AN Zhixing, LI Yu, et al. Cloned goats (Capra hircus) from adult ear cells. SCIENCE IN CHINA (Series C).2002,45(3):260-267
[227]刘凤军,工勇胜,华松。培养液及血清浓度对山羊孤雌胚胎体外培养的影响。黑龙江畜牧兽医,2004,2：11-13
[2]Vollrath F. Strength and structure of spider'silks. Biological Macromolecules,2000,74:67-83
[3]何兰芝,陈莉萍,王学梅。二十一世纪新型纤维——蜘蛛丝。甘肃科技,2004,20(11)：133～134
[4]S.F.Y.Li et al.Comparative Study of the Internal Structure of Kelvar and Spider Silk by Atomic Force Microscopy.J.Vac.Sci.Technol.A.,1994(4):1891-1894
[5]S.F.Y.Li et al.New Internal Structure of Spider Silk Revealed by Atomic Force Microscopy. Biophysical Journal,1994 (66):1209-1212
[6]Kitagawa et al.Microstructure of Spider Dragline.J.Soc.Mat.Sci.,2001(50):1213-1217
[7]Augsten K et al. Glycoproteins and Skin-core Structure in Nephlia Clavipes Spider Silk Observed by Light and Electron.Scanning,2000(1):12-15
[8]S.Frische et al.Elongate Cavities and Skin-core Structure in Nephlia Spider Silk Observed by Electron Microscopy. J. Microscopy,1998(189):64-74
[9]Z.Shao et al.Heterogeneous Morphology of Nephila Edulis Spider Silk and its Significance for Mechanical Properties. Polymer,1999 (40):4709-4711
[10]Sirichaisit J, Yong R J, Vollrath F. Molecular deformation in spider dragline silk subjected to stress. Polymers,1999,40:2493-2500
[11]Riekel C, Muller M, Vollrath F. In-situ X-ray diggraction studies during forced silking of a Nephila senigalensis spider. Macromocules,1999,32:4464-4466
[12]Hijirida H, Do K G, Michal, et al. C-13 NMR of Nephial clavipes major ampulllate silk gland. Biophys,1996,71:3442-3447
[13]Shao Z Z, Vollrath F, Sirichaisit J. Analysis of spider silk in native and supercontracted states using Raman spectroscopy. Polymer,1999,40:2493-2500
[14]刘海洋,工伟霞,刘长军等。纺织新材料——蜘蛛丝。纺织导报,2004(1)：28—30
[15]周文聪,张贤,张敏等。影响蜘蛛丝性能的因素及开发技术。纺织科技进展,2010(4)：58-61
[16]赵博。新一代高性能纤维—蜘蛛丝纤维。国外丝绸,2005,(1)：22-23
[17]刘庆生,段亚峰。蜘蛛丝的结构性能与研究现状。四川丝绸,2005,2：16-18
[18]袁辉。蜘蛛丝的研究与开发利用。国外纺织技术,2006,6：7-9
[19]蒋平,肖永红,吕太勇等。蜘蛛丝的结构与机械性能研究进展。井冈山大学学报,2010,31(4)：37-43
[20]陈瑶,孟清,卿凤翎。蜘蛛丝纤维的特性与开发生产。现代纺织技术,2006,6：53-56
[21]J. Gatesyetal. Extreme diversity conservation and convergence of spider silk fibroin sequences. Science,2001,291:2603-2605
[22]段亚峰,冀勇斌。蜘蛛丝开发应用的现状与进展。丝绸,2002,7：46-47
[23]王琳纲,胡平。蜘蛛丝的研究进展及展望。新材料产业,2007,9：63～67
[24]J. Gatesyetal. Extreme diversity, Conservation, and convergence of spider silk fibroin sequences. Science,2001,291:2603-2605
[25]F. Vollrath and D. P. Knight. Liquid Crystalline Soinning of Spidersilk. Nature,2001,410: 545-548
[26]黄献聪,施楣梧。蜘蛛丝的力学性能及其应用取向。纺织导报,2004,3：32-38
[27]潘志娟,邱芯薇。蜘蛛丝的物理性能研究。苏州大学学报,2003：23(1)：18-22
[28]刘辉芬,蒋平,肖永红。两种不同功能蜘蛛丝的物理化学结构表征与力学性。武汉大学学报,2010,56(1)：93-99
[29]盛家墉,潘志娟,陈宇岳。蜘蛛丝的化学组成与结构初探,2000,4：8-13
[30]刘全勇,江雷。仿生学与天然蜘蛛丝仿生材料。高等学校化学学报,2010,31(6)：1065-1071
[31]张慧勤,王志新。蜘蛛丝的研究与应用。中原工学院学报,2005,16(4)：47-50
[32]Xu M, Lewis R V. Structure of a protein superfiber:spider dragline silk. Proc Natl Acad Sci U S A,1990,87(18):7120-7124
[33]Hinman M B, Lewis R V. Isolation of a clone encoding a second dragline silk fibroin. J Biol Chem,1992,267 (27):19320-19324
[34]Colgin M A, Lewis R V. Spider minor ampullate silk proteins contain new repetitive sequences and highly conserved non-silk-like "spacer regions". Protein Sci,1998, 7(3):667-672
[35]Beckwitt R,Arcidiacono S.Sequecn conservation in the C-teminal region of spider silk proteins(Spidroin) from Nehpila clavipes(Tetragnathidae) and Araneus bicentenarius(Araneidae).J.Biol.Chem; 1994,269:6661-6663
[36]Hayashi C Y, Lewis R V. Evidence from flagelliform silk cDNA for the structural basis of elasticity and modular nature of spider silk. J Mol Biol,1998,275(5):773-784
[37]Guerette P,Ginzinger D,Weber B,Gosline J. Silk properties determined by gland-specific expression of a spider fibroin gene family.Science,1996.272:112-115
[38]Prince J T, Mcgrath K P, DiGirolamo C M, et al. Construction, cloning and expression of synthetic genes encoding spider dragline silk. Biochemistry,1995, 34(34):10879-10885
[39]Xia X X, Qian Z G, Ki C S, et al. Native-sized recombinant spider silk protein produced in metabolically engineered Escherichia coli results in a strong fiber. Proc Natl Acad Sci U S A,2010,107(32):14059-14063
[40]Fahnestock R, Bedzyk L A. Production of synthetic spider dragline silk protein in picha pastoris. Appl Microbiol Biothechnol,1997,47(1):33-39
[41]Scheller J, Guhrs K H, Grosse F, et al. Production of spider silk proteins in tobacco and potato. Nat Biotechnol,2001,19(6):573-577
[42]Scheller J, Henggeler D, Viviani A, et al. Purification of spider silk elastin from transgenic plants and application for human chondrocyte proliferation. Transgenic Res, 2004,13(1):51-57
[43]Piruzian E S, Bogush V G, Sidoruk K V, et al. Construction of synthetic genes for analogs of spider silk spidroin 1 and their expression in tobacco plants. Mol Biol, 2003,37(4):554-560
[44]Barr L A, Fahnestock S R, Yang JJ, et al. Production and purification of recombinant DP1B silk-like protein in plants. Mol Breeding,2004,13(4):345-356
[45]Lazaris A, Arcidiacono S, Huang Y, et al. Spider silk fiber spun from soluble recombinant silk produced in mammalian cells. Science,2002,295(5554):472-476
[46]Gatesy J, Hayashi C, Motriuk D, et al. Extreme diversity, conservation, and convergence of spider silk fibroin sequences. Science,2001,291(5513):2603-260519324
[47]Xu H T, Fan B L, Yu S Y, et al. Construct synthetic gene encoding artificial spider dragline silk protein and its expression in milk of transgenic mice. Anim Biotechnol, 2007,18(1):1-12
[48]Ponder BA. Cancer genetics. Nature.2001.411(6835):336-341
[49]梁洋。蜘蛛拖丝蛋白基因Spidroinl嵌合体小鼠的研究。东北林业大学博士论文.2011,6
[50]Carl A, Pinkert. Transgenic Animal Technology:A Laboratory Handbook. Second Edition. Chenical Industry Press,2004,134-135
[51]Buchschacher Jr GL.Introduction to retroviruses and retroviral vectors. Somat Cell Mol Genet,2001,26:1-11
[52]Hu W S and Pathak V K. Design of Retroviral Vectors and Helper Cells for Gene Therapy. Pharmacol Rev Dec,2000,52(4):493-511
[53]唐东红,代解杰,仝品芬等。泡沫病毒载体的研究进展。病毒学报,2000,9(3)：282-284
[54]Yu S F, Sullivan M D, Linial M L. Evidence that the human foamy virus genome is DNA. J Virol,1999,73:1565-1572
[55]许建,李世崇,陈昭烈。逆转录病毒表达系统及其在外源蛋白高效表达中的应用。 中国生物工程杂志,2008,28(5)：116-121
[56]王振发,王烈,卫立辛。基因治疗病毒载体的研究进展,2007,4(3)：490-492
[57]Dalba C, Bellier B, Kasahara N et al..Replication-competent Vectors and Empty Virus-like Particles:New Retroviral Vector Designs for Cancer Gene Therapy or Vaccines.The American Society of Gene. Therapy Mol Ther,2007,15(3):457-466
[58]Yu S S, Han E, Iiong Y et al. Construction of a retroviral vector production system with the minimum possibility of a homologous recombination. Gene Ther,2003,10:706-711
[59]Dillon P, Lenz J, Rosen C. Construction of a replication-competent murine retrovirus vector expressing the human immunodeficiency virus type 1 tat transactivator protein. J Virol,1991,65:4490-4493
[60]Solly S, Trajcevski S, Frisen C et al. Replicative retroviral vectors for cancer gene therapy. Cancer. Gene Ther,2003,10:30-39
[61]Wang W, Tai C, Kasahara N et al. Highly efficient and tumor-restricted gene transfer to malignant gliomas by replication-competent retroviral vectors. Hum Gene Ther,2003,14:117-127
[62]Trajcevski S, Solly S, Frisen C et al. Characterization of a semi-replicative gene delivery system allowing propagation of complementary defective retroviral vectors. J Gene Med,2005,7:276-287
[63]Doan L, Li M, Chen C et al. Virus-like particles as HIV-1 vaccines. Rev Med Virol,2005,15:75-78
[64]Robertson E, Bradley A, Kuehn M, et al. Germ-line transmission of genes introduced into cultured pluripotential cells by retroviral vector. Nature,1986,323(6087):445-448
[65]Chan A W. Homan E J, Ballou L U et al. Transgenic cattle produced by reverse-transcribed gene transfer in oocytes. Proc Natl Acad Sci USA,1998,95(24):14028-14033
[66]Chan A W, Chong K Y, Martinovich C et al. Transgenic Monkeys Producedby Retroviral Gene Transferinto Mature Oocytes. Science,2001,291(5502):309-312
[67]Matsunar H, Onodera M, Tada N et al. Transgenic-Cloned Pigs Systemically Expressing Red Fluorescent Protein, Kusabira-Orange. Cloning Stem Cells,2008,10(3):313-323
[68]Benigni A and Remuzzi G. Gene Therapy for Renal Diseases and Transplantation. Contrib Nephrol Basel Karger,2008,159:30-46
[69]Rainov N G, Kramm C M, Banning U et al. Immune response induced by retrovirus-mediated HSV-tk/GCV pharmacogene therapy in patients with glioblastoma multiforme. Gene Ther,2000,7(21):1853-1858
[70]谢文练,黄健,韩金利等。逆转录病毒介导HSV-TK基因结合更昔洛韦对裸鼠膀胱癌模型的抑癌作用。中华泌尿外科杂志,2005,26(11)：752-754
[71]Hauster H,Gross G.Brunsw, et al. Inducibility of humanB-Interferon gene in mouse L-cell clones. Nature,1982,297:650
[72]孔永,秦秀云。动物细胞培养技术研究进展。重庆文理学院学报,2007,26(4)：54-56
[73]谢文忠。巯嘌吟键连卟啉抗癌药物的合成、表征及生物活性研究。湖南大学硕士学位论文,2008,19
[74]肖深初。三嗪卟啉衍生物的合成、表征及其生物活性研究。湖南大学博士学位论文,2009,63
[75]魏宗波。雏鸡心肌细胞原代培养及锰对肉仔鸡心肌细胞MnSOD基因表达调控研究。西北农林科技大学,2008,3
[76]冯为慧。淡水鱼类细胞现采现培技术的初步建立与关键技术点研究。南昌大学硕士学位论文,2012,10
[77]顾涵英,冯佑民。无血清细胞培养基及其主要补充因子。生物化学与生物物理进展,1990,17(4)：255-259
[78]马忠仁,冯玉萍,李倬。动物血清在细胞培养中的重要性及其质量控制标准。西北民族大学学报(自然科学版)。2003,24(50)：56-58
[79]刘艳。细胞培养及常见问题的处理。山东畜牧兽医,2008,29：13-16
[80]甘露,张志辉,吕美娜。细胞培养技术。中国组织工程研究与临床康复,2008,12(29)：5739-5742
[81]王立新,杨朝霞。动物细胞培养及应用。黄牛杂志,2000,26(3)：45-48
[82]桂东城,蒋涛。转基因克隆动物的研究进展。中国畜牧兽医,2009,36(11)：114-117
[83]骆继怀,独军政,常惠芸。转基因克隆动物的发展及其应用。动物医学进展,2010,31(7)：103-106
[84]刘西梅,乔宪凤,张立苹等。转基因动物研究的技术思路。湖北农业科学,2012,51(21)：4837-4840
[85]Gordon J W,Scangos G A,Plotrin D J,et al.Genetic transformation of mouse embryo by microinjection of purified DNA.Proc Natl A-cad Sci USA,1980,77:7380-7384
[86]余华,叶健强,刘丹。动物转基因及克隆技术研究。信阳农业高等专科学校学报,2009,19(2)：104-108
[87]刘晓兰,王仁华,工峥等。动物转基因技术及其应用。湖南饲料,2010,1：28-31
[88]Evans M J,Kaufman M H. Establishment in culture of pluripotential cells from mouse embryos. Nature,1981,292:154-156
[89]刘黎青,李娟,周盛年。胚胎干细胞及在生殖医学中的应用。中国组织工程研究与临床康复,2011,15(1)：113-115
[90]张瑞杰,苗向阳。动物转基因技术研究现状与应用前景。中国细胞生物学报,2010,32(5)：800-807
[91]王玉琴。绵羊同期发情研究进展。畜禽业,2003,4：4-6
[92]蒋长炜,冯国献。绵羊同期发情技术概述。畜牧与饲料科学。2010,31(10)：109-110
[93]李武,王丽娟,李丽华。绵羊胚胎移植研究进展。黑龙江畜牧兽医,2002,7：48-50
[94]傅文栋,李风德,牛建克。绵羊胚胎移植研究进展。黑龙江动物繁殖。2010,18(5)：20-21
[95]Gordon JB, Uehlinger V.Fertile intestine nuclei.Nature.1966,210:1240-1241
[96]McGrath J and Sloter D.Inability of mouse blastomere nuclei transtected to enucleated zygotes to support development in vitro.Science,1984,226:1317-1319
[97]朵红。动物核移植技术研究概况。2005,35(4)：43～44
[98]孙志宏,邱瑞,李延清等。哺乳动物核移植研究。安徽农业科学,2008,36(7)：2783-2784,2788
[99]Wilmut I, Schnieke A E, McWhir J, et al. Viable offspring derived from fetal and adult mammalian cells. Nature,1997,385:810-813
[100]Baguisi A, Behboodi E, Melican D T, et al. Production of goats by somatic cell nuclear transfer. Nat Biotechnol,1999,17 (5):456-461
[101]Kubota C, Yamakuchi H, Todoroki J, et al. Six cloned calves produced from adult fibroblast cells after long-term culture. Proc Natl Acad Sci,2000,97(3):990-995
[102]Dai Y, Vaught T D, Boone J, et al. Targeted disruption of the alphal,3-galactosyl transferase gene in cloned pigs Nat Biotecnol,2002,20(3):251-255
[103]Wakayama T, Rodriguez I, Perry A C, et al. Mice cloned from embryonic stem cells. Proc Natl Acad Sci USA..1999,96(26):14984-14989
[104]Chesne P, Adenot P G, Viglietta C, et al. Cloned rabbits produced by nuclear transfer from adult somatic cells. Nat Biotechnol,2002,20(4):366-369
[105]Shin T, Kraemer D, Pry or J, et al. A cat cloned by nuclear transplantation. Nature, 2002,415 (6874):859
[106]Lagutina I, Lazzari G, Duchi R, et al. Somatic cell nuclear transfer in horses:effect of oocyte morphology, embryo reconstruction method and donor cell type. Reproduction,2005,130(4):559-567
[107]杨彩侠,陈大元。动物克隆机理研究进展。动物学研究,2003,24(1)：73-78
[108]Kubota C, Yamakuchi H, Todoroki J, et al. Six cloned calves produced from adult fibroblast cells after long-term culture.Proc Nat Acad Sci USA,2000,97(3):990-995
[109]Enright B P, Jeong B S, Yanget X, et al. Epigenetic characteristics of bovine donor cells for nuclear transfer:levelsof histone acetylation.Biology of Reproduction,2003,69(5):1525-1530
[110]Poehland R, Al-Rostum F, Becker F, et al. Donor cell lines considerably affect the outcome of somatic nuclear transfer in the case of bovines. Journal of Reproduction and Development,2007,53 (4):737-748
[111]Betthauser J, Forsberg E, Augenstein M, et al. Production of cloned pigs from in vitro systems. Nat Biotechnol,2000,18(10):1055-1059
[112]Hill J R, Winger Q A, Burghardt R C, et al. Bovine nuclear transfer embryo development using cells derived from a cloned fetus. Anim Reprod Sci,2001,67(1 12):17-26
[113]Takahashi K, Yamanaka S. Induction of pluripotent stem cells frommouse embryonic and adult fibroblast cultures by defined factors. Cell,2006,126(4):663-679
[114]Schwarzer M, Carnwath J W, Lucas-Hahn A, et al. Isolation of bovine cardiomyocytes for reprogramming studies based on nuclear transfer. Cloning Stem Cells,2006,8(3):150-158
[115]Mizutani E, Ono T, Li C, et al. Propagation of senescent mice using nuclear transfer embryonic stem cell lines. Genesis,2008,46(9):478-483
[116]张传山,郭毅,谷瑞环等。成体兔转基因成纤维细胞的克隆分离及其核移植研究。生物化学与生物物理进展,2009(9)：1186-1192
[117]Tao Y, Han W, Zhang M, et al. Production of Boer goat (Capra hircus) by nuclear transfer of cultured and cryopreserved fibroblast cells into slaughterhouse derived oocytes. Czech Journal of Animal Science,2009,54 (10):448-460
[118]工建锋,张勇,赵兴绪。不同采卵方式对绵羊卵母细胞质量、体外成熟及体外受精的影响。动物医学进展,2007,28(8)：51-54
[119]万鹏程,张云生,石文艳。绵羊卵母细胞不同采集方法对体外成熟培养的影响。石河子大学学报,2004,22(5)：401-404
[120]Keskintepel L, Darwish G M, Kenimer A T, et al.Term development of caprine embryos derived from immature oocytes in vitro.Theriogenology,1994,42(3):527-535
[121]许杰,马恒东,吴兴杰。采集方法、EGF对山羊卵母细胞体外成熟的影响。安徽农业科学,2007,35(17)：5185-5186,5191
[122]刘青春,李武峰,岳文斌等。不同卵巢运输温度和培养方法对卵母细胞体外成熟的影响。畜牧兽医杂志,2008,27(4)：16-17
[123]汪立芹,杨梅,陈童等。季节和方法对牛、绵羊卵母细胞回收及体外成熟的影响。中国畜牧兽医,2008,35(11)：74-76
[124]张伟,工利红。影响卵母细胞体外成熟的因素。黄牛杂志,2005,31(1)：51-54
[125]Mayes MA, Sirard MA. The influence of cumulus oocyte complex morphology and meiotic inhibitors on the kinetics of nuclear maturation cattle. Theriogenology, 2000,54:911-922
[126]Masui Y and markert C L.Cytoplasmic centrol of nuclear behavior during meiotic muturation of frog oocytes.J Exp Zool,1971,177:129-146
[127]Smith L D, Ecker.The interaction of steroids with Rena pipiensoocytes in the induction of maturation.Dev Biol,1971,25:232-247
[128]Wakayama T, Perry A C F, Zoccotti M, et al.Full-tem development of mice from enucleated oocytes injected with cumulus cell nuclei.Nature,1998,394 (6691):369-374
[129]Zhou Q, Jouneau A, Brochard V, et al.Development potential of mouse embryos reconstructed from metaphase embryonic stem cell nuclei.Biol Reprod,2001,65 (2):412-419
[130]Lanza R P, Cibelli J B, Diaz C, et al.Cloning of endangered species (Bos Gaurus) using interspecies nuclear transfer.Bio.Reprod.,2001,64:1487-1493
[131]Kono T, Obata Y, Wu Q, et al. Birth of parthenogenetic mice that can develop to adulthood.Nature,2004,428 (6985):860-864
[132]李清。卵母细胞去核方法及其对卵母细胞发育的影响。山东畜牧兽医,2007,28(6)：42-44
[133]Wilmut I, Schnieke A E,McWhir J,et al. Viable offspring derives from fetal and adult mammalian cells.Nature,1997,385:810-813
[134]Illmensee K,Hoppe PC.Nuclear transplantationin mouse musculus.development potential of nuclei from preimplantation embryos.Cell,1981,23:9-16
[135]Willadsen SM. Nuclear transplantation in sheep embryo. Nature,1986,320:63-65
[136]Prather RS, Barnes FL, Sims MM, et al.Nuclear transplantation in the bovine embryos: Assessment of donor nuclei and recipient oocyte. Biol Reprod,1987,37:859-866
[137]Stice SL, Robl JM. Nuclear reprogramming in nuclear transplant rabbit embryo. Biol Repod,1988,39:657-664
[138]Prather RS, Sims MM, First NL. Nuclear transplantation in early pig embryos. Biol Repod,1989,44:414-418
[139]杜淼,陆德裕,沈三兵。家兔早期胚胎细胞发育能力的研究。实验生物学报,1990,23(4)：477-482
[140]张涌,王建辰,钱菊汾等。山羊卵核移植的研究。中国农业科学,1991,24(5)：1-6
[141]邹贤刚,李光三,王玉阁。山羊胚胎细胞经继代细胞核移植后其发育能力的研究。科学通报,1995,40(3)：264-267
[142]李雪峰,石德顺,谭丽玲等。电激活对完全体外化牛细胞核移植的影响。畜牧兽医学报,1996,27：495-500
[143]赵浩斌,陈乃清,魏庆信等。猪卵核移植的研究。武汉大学学报,1997,43 (4)：505-510
[144]Li M, John J, Richard L, et al. Rhesus monkey produced by nuclear transfer [J]. Biol Reprod,1997,57:454-459
[145]Wakayama T,Perry ACF, Zoccotti M et al. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature,1998,394(7):369-374
[146]Cibelli J B,Stice S L,Golueke P J,et al. Cloned transgenic calves produced from nonquiescent fetal fibroblasts.Science,1998,280:1256-1258
[147]132.Vignon X,Chesns P,LebourhisD.Dvelopmental Potential of bovine embryos reconstructed with somatic nuclei from cultured skin and muscle fetal cells.Theriogenology,1998,49:392
[148]KatoY,Tani T,Sotomaru Y,etal.Eight calves cloned from somatic cells of a single adult.Science,1998,282:2095-2098
[149]Well D N,Misica P M,Tervit H R,etal.Production of cloned calves following nuclear transfer with cultured adult mural granulose cells.Biology of ReProduetion,1999,60:996-1005
[150]Baguisi A, Behboodi E, Melican DT, Pollock JS, Destrempes MM, Cammuso C, Williams JL, Nims SD, Porter CA, Midura P, Palacios MJ, Ayres SL, Denniston RS, Hayes ML, Ziomek CA, Meade HM, Godke RA, Gavin WG, Overstrom EW,Echelard Y. Production of goats by somatic cell nuclear transfer.Nat Biotechnol,1999;17(5):456-461
[151]工玉阁,邹贤刚,成国祥等。由胎儿成纤维细胞而来的克隆山羊。科学通报,1999,44(21)：2319-2323
[152]OnishiA,Iwamoto M,Akita J,etal.pigs cloning by microinjeetion of fetal fibroblast nuclei.Science.2000,289:1187-1193
[153]McCreath K J, Howcroft J, Campbell K H,et al.Production of gene-targeted sheep by nucleartransfer from cultured somatic cells.Nature,2000,405:1066-1069
[154]Chesne P, Adenot P G, Viglietta,et al. Cloned rabbits produced by nuclear transfer from adult somatic cells.Nat Biotechnol,2002,20(4):366-369
[155]Li, Z. Sun, X. Chen, J.etal.Cloned ferrets produced by somatic cell nuclear transfer. Dev Biol.2006. (293):2
[156]Nisar A.Wani, U.Wernery,F.A.H.Hassan,4R.et al.Production of the First Cloned Camel by Somatic Cell Nuclear Transfe. Biol Reprod,2010,82:373-379
[157]Briggs R,King T J.Transplantation of living cell nuclei from blastula cells into enucleated frogps eggs.Proc Natl Acad Sci USA,1952,38:455-463
[158]童第周,吴尚勤,叶毓芬等。鱼类细胞核的移植。科学通报,1963,7：60-61
[159]童第周,叶毓芬,陆德裕等。鱼类不同亚科间的细胞核移植。动物学报,1973,19(3)：201-209
[160]McGrath JD,etal.Inability of mouse blastomere nuclei transferred to enucleated zyzotes tosupport development in vitro.Scienee,1984,226:1317
[161]WolfeBA,etal.Methods in bovine nuclear transfer.Theriogenology,1992,37:5
[162]梅祺,邹贤刚,杜森。鼠兔核质杂交胚胎早期发育的研究。实验生物学报,1993：26(4)：389-397
[163]陈大元,孙青原,刘冀珑等。大熊猫供核体细胞在兔卵胞质中可去分化而支持早期重构胚的发育。中国科学(C辑),1999,29(3)：324-330
[164]Dominko T,Mitalipova M,Haley B,et al.Bovine oocytes cytoplasm supports development of embryos produced by nuclear transfer of somatic cell nuclei from various mammalian species.Biology of Reproduction,1999,60:1496-1502
[165]Lanza RP, Cibelli JB, Diaz F, et al, Cloning of an endangered species (Bos gaurus) using interspecies nuclear transfer. Cloning 2000,2(2):79-90
[166]Murakami M, Otoi T, Wongsrikeao P, et al.Development of interspecies cloned embryos in yak and dog.Cloning Stem Cells,2005,7(2):77-81
[167]Thongphakdee A, Numchaisrika P, Omsongkram S, et al.In vitrodevelopment of marbled cat embryos derived from inter-species somatic cell nuclear transfer. Reprod Domest Anim,2006,41 (3):219-226
[168]Sugimura S., Narita K., Yamashiro H., Sugawara A., Shoji T.,Terashita Y., Nishimori K., Konno T., Yoshida M., and Sato E., Interspecies somatic cell nucleus transfer with porcine oocytes as recipients:A novel bioassay system for assessing the competence of canine somatic cells to develop into embryos, Theriogenology,2009,72(4):549-559
[169]Sha H.Y., Chen J.Q., Chen J., Zhang P.Y., Wang P., Chen L.P.,Cheng GX., and Zhu J.H., Fates of donor and recipient mitochondrial DNA during generation of interspecies SCNT-derived human ES-like cells, Cloning and Stem Cells,2009,11(4):497-507
[170]舒金辉,陶锋,张波。体细胞核移植表观遗传的研究进展。中国比较医学杂志,2008,18(8)：49-52
[171]刘秀丽。哺乳动物核移植技术研究进展。贵州畜牧兽医,2006,30(2)：8-11
[172]周艳华,贾银海,苏雷等。影响细胞核移植效率的因素。现代畜牧兽医,2008,2：46-50
[173]王建,向舒,陆凤花等。哺乳动物体细胞核移植后核重编程的研究进展。安徽农业科学,2010,38(25)：13732-13733
[174]汪立芹,王静,赵云程等。供核细胞及处理方式对绵羊核移植效率的影响。华北农学报,2012,27：69-73
[175]王春生。转拟蜘蛛拖丝蛋白基因绵羊成纤维细胞系的建立。东北农业大学博士论文。2007,55
[176]原璐。转拟蜘蛛拖丝蛋白基因四聚体绵羊成纤维细胞株的筛选。东北林业大学硕 士论文。2009,20
[177]薛庆善。体外培养的原理与技术,北京：科学出版社,2001,2-6
[178]Kazutoshi Takahashi, Shinya Yamanaka. Induction of Pluripotent Stem Cells from Mouse Embryonic and Adult Fibroblast Cultures by Defined Factors. Cell,2006,126:663-676
[179]Haslam S Z. Progesterone effects on deoxyribonucleic acid synthesis in normal mouse mammary glands. Endocrinology.1988,122(2):464-470
[180]李扬,吴凯峰,郭旭东等。脂质体介导外源基因体外转染牛胎儿成纤维细胞条件的优化。遗传,2002,24(6)：653-655
[181]佟慧丽。奶山羊乳腺上皮细胞系的建立。东北农业大学硕士论文,2007,11
[182]刘亚,章孝荣,张运海等。牛、羊成纤维细胞的冷冻与冷藏保存。中国兽医学报,2004,24(2)：197-198
[183]翟秀岩,马文锋,热海佐保子。培养细胞的冷冻保存和解冻复苏的操作方法。中国医科大学学报,2002,31(1)：77-78
[184]王晓玉,柳增善,任洪林。“生物刚性及弹性蛋白”—蜘蛛丝研究进展。第四军医大学学报,23：31-34
[185]Hinman, M B, Jones, J.A.&Lewis R V. Synthetic spider silk:a modular fibre. TIBTECH, 2000,18:374-379
[186]Scheller J, Guhrs KH, Grosse F et al. Production of spider silk proteins in tobacco and potato. Nat Biotechnol,2001,19:573-577
[187]Pelletier J, Flynn M E, Kaplan G, Racaniello V, Sonenberg N. Mutational analysis of upstream AUG codons of poliovirus RNA. J Virol.1988,62(12):4486-4492
[188]郑月茂。转基因山羊乳腺上皮细胞系建立与转基因克隆胚发育。西北农林科技大学博士论文,2005,132
[189]王沛涛,何立群,舒志全等。细胞低温生物学研究进展。齐鲁医学杂志,2005,20：1-3
[190]潘琼,陈彦,梅黎等。冷冻复苏对大鼠干细胞代谢活性的影响。中国药理学与毒理学杂志,2008,22(6)：457-462
[191]杜晨光。绵羊卵母细胞孤雌激活与体细胞核移植的研究。内蒙古大学硕士学位论文,2005,7
[192]孙杰。影响绵羊卵丘细胞核移植效果主要因素的研究。河北农业大学硕士学位论文,2008,17.
[193]赖彦。绵羊核移植技术的研究。内蒙古农业大学硕士学位论文,2010,18
[194]南木甲,张雁平。不同采卵方法及卵巢所处性周期阶段对绵羊卵母细胞体外成熟的影响。中国畜牧兽医,2007,34(7)：137-139
[195]张永胜,工宏伟,皮文辉。两种采卵方法对绵羊卵母细胞体外受精的影响。甘肃 畜牧兽医,2005,3：9-11
[196]Sommersberg B,Bulling A,SalzerU,et al.Gap junction communication and connexin 43 gene expression in a rat granulose cell line:regulation by follicle-stimulating hormone.BioReprod,2000,63(6):1661-1668
[197]周平,刘东军,闫真等。TGFα、EGF对绵羊卵母细胞成熟的影响。细胞生物学杂志,2007,29：713-717
[198]郭延华。EGF对不同直径卵泡绵羊卵母细胞体外成熟、受精及胚胎发育的影响。石河子大学硕士学位论文,2010,2
[199]毛晓霞,万双秀。生殖激素对绵羊卵母细胞体外成熟的影响。上海畜牧兽医通讯。2009,1：33-34
[200]张晓建,安志兴,李学斌。卵母细胞体外成熟时间对绵羊核移植效率的影响。安徽农业科学,2008,36(32)：14120-14121,14132
[201]Albertini D F, Combelles C M, Benecchi E, et al. Cellular basis for paracrine regulation of ovarian follicle development。Reproduction,2001,121,647-653
[202]赛务加甫。提高绵羊体细胞核移植效率的研究。西北农林科技大学博士论文,2007,41
[203]曲杨燕,赵雁伟,李凯等。FSH对绵羊卵母细胞体外核成熟的影响。中国畜牧兽医,2008,35(12)：80-83
[204]郭宪,陈学进,叶荣等。牛卵泡卵母细胞体外成熟的研究。中国草食动物,2004,24(2)：3-6
[205]林雍峰,赵兴绪,张勇。绵羊卵母细胞核成熟、卵丘扩展与激素浓度关系的研究。甘肃农业大学学报,2012,47(1)：22-27
[206]梁素丽,李向臣,白修云等。影响卵母细胞体外成熟的因素。中国畜牧兽医,2007,34(9)：36-39
[207]何良军,彭莉,马兵祥等。绵羊卵母细胞最优运输温度和体外成熟时间的研究。塔里木大学学报,2005,17(4)：5-7
[208]乔利敏,郭彤,乔富强。绵羊卵巢运输、保存温度及时间对其卵母细胞孤雌发育能力的影响。浙江农业学报,2012,24(4)：563—566
[209]王旭光。绵羊卵母细胞体外成熟和孤雌激活的研究。新疆农业大学硕士论文。2003,27
[210]王建辰,章孝荣。动物生殖调控。安徽科学技术出版社,1998：220～230
[211]Machaty Z, PratherR S. Strategies for activating nuclear transfer oocytes. Reprod Fertil Dev,1998,10:599-613
[212]Tateno H, Kamiguchi Y.1997. Parthenogenetic activation of Chinese hamster oocytes by chemical stimuli and its cytogenetic evaluation. Mol Reprod Dev.47:7278
[213]Morgan A and Jacob R. Ionomycin enhances influx by stimulating store-regulated cation entry not by a direct action at the plasma membrane. Bio Chem J.1994,300:665 672
[214]Wang WH, Machaty Z, Ruddoch N, et al.Activation of porcine oocytes with alcium ionophore:effects of extracellularcalcium.Mol Reprod,1999 (1):99-107
[215]施巧婷,王旭光,王立虎。中国畜牧杂志。2004,40(6)：10-13
[216]张晓建。绵羊卵母细胞孤雌激活与体细胞核移植。西北农林科技大学硕士论文,2005,30
[217]彭礼繁,罗光彬。牛、羊卵母细胞孤雌激活的研究。畜禽业,2008,226：56-58
[218]周欢敏,李宝山等。不同激活方法对绵羊体外成熟卵母细胞孤雌生殖的影响。内蒙古农业大学学报,2008,29(1)：78-81
[219]张云生。绵羊卵母细胞孤雌激活及冷冻研究。石河子大学硕士论文。2005,22
[220]陆凤花,舒金辉,黎江等。哺乳动物克隆电融合技术的影响因素。广西农业生物科学,2007,26(2)：179-182
[221]王立虎,黄俊成,杨梅。绵羊体细胞核移植实验研究。草食家畜,2001,111：17-19
[222]工亮,彭涛,朱海等。电融合条件对新疆盘羊(Ovis ammon)和转基因(GFP)萨能奶山羊核移植胚胎发育的影响。西北农业学报,2006,15(2)：1-6
[223]H.R.TERVIT,D.G.WHITTINGHAM and L.E.A.ROWSON.Successful culture in vitro of sheep and cattle ova.J.Reprod.Fert.1972,30,493-497
[224]Grazyna Ptak,Pasqualino Loi,Maria Dattena,et al.Offspring from One-Month-Old Lambs: Studies on the Developmental Capability of Prepubertal Oocytes. Biology of reproduction,1999,61:1568-1574
[225]Pasqualino Loi,Michael Clinton,Barbara Barboni, et al.Nuclei of Nonviable Ovine Somatic Cells Develop into Lambs after Nuclear Transplantation. Biology of reproduction, 2002:67,126-132
[226]GUO Jitong, AN Zhixing, LI Yu, et al. Cloned goats (Capra hircus) from adult ear cells. SCIENCE IN CHINA (Series C).2002,45(3):260-267
[227]刘凤军,工勇胜,华松。培养液及血清浓度对山羊孤雌胚胎体外培养的影响。黑龙江畜牧兽医,2004,2：11-13
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |