草菇开伞基因克隆及反义表达载体构建
|
摘要
草菇子实体的发育大致经过以下六个阶段：针头期(Pinhead)、小纽扣期(Tiny button)、纽扣期(Button)、蛋形期(Egg)、伸长期(Elongation)和成熟期(Mature)。子实体在不同时期的生理特征变化显著，尤其是在蛋形期进入伸长期时呼吸强度开始急剧增强，子实体的形态也从未破膜的卵形变成开伞，失去商品价值。是什么原因使草菇子实体在短时间内产生这么大的变化?本研究通过细胞形态变化，激素水平变化以及特异基因的表达等方面进行了探索研究。主要研究及结果如下：
1．细胞伸长主导伸长期子实体菌柄快速伸长 显微观察草姑子实体菌柄细胞在不同发育时期以及菌柄不同部位在蛋形期和伸长期的细胞形态并测量了细胞大小变化，同时通过菌柄标记观察并测量了子实体菌柄的伸长区间。结果表明：在蛋形期草菇主要是通过细胞分裂使细胞数增加而使子实体长大，从伸长期开始，子实体菌柄伸长的主导因素是菌柄中上部细胞的伸长，同时菌柄顶部细胞的分裂起辅助作用。
2．内源激素水平变化与子实体生长显著相关 测量子实体菌柄在不同发育阶段的内源激素水平变化，结果显示：在蛋形期之前，无论促进生长类激素还是衰老类激素都没有明显的变化；但是在伸长期，促进生长类激素GA_(1/3)和ZRs急剧上升到顶峰，衰老类激素ABA则急剧下降；到成熟期，GA_(1/3)和ZRs的含量比伸长期时明显下降，但仍维持在较高的水平，ABA的浓度在成熟期时下降到最低水平。在不同发育时期中GA_(1/3)的浓度变化主导GA_(1/3)／ABA的比率变化，GA_(1/3)／ABA可能是主导菌柄细胞生长的重要因素，但子实体进入成熟期后的衰老过程可能有其它控制衰老的生理因子起关键作用。
3．克隆到开伞阶段特异表达基因片段 通过抑制性扣除杂交法，以蛋形期子实体菌柄的Total mRNA为驱赶子，伸长期子实体菌柄的Total mRNA为目的子，克隆子实体菌柄在伸长期特异表达的基因，结果得到一个基因片段，测序并登录GENBANK进行同源性比较，没有与此序列相同的基因。以该基因片段为模板合成引物并进行基因组DNA扩增，双引物扩增到目的片段，单引物扩增得到1～2KB左右的扩增片段。
4．正、反义基因片段表达载体的构建 酶切分离克隆质粒中的目的基因片段，同时对酶切的目的表达载体进行末端平滑及去磷酸化处理，通过平末端连接，将克隆的基因片段插入到PCB载体的多克隆位点中，转化大肠杆菌，随机挑选阳性菌落，提取质粒进行测序，证实获得分别具有正反序列的克隆载体。
The development of Volvariella volvacea fruit-body has six stages of the pinhead,the tinny button,the button,the egg,the elongation and the mature.The physiological characteristics of the fruit-body changes obviously in different stages, respiratiry intensity enhanced dramatically especially from the egg stage to the elongation stage.At the same time , the morpha of the fruit-body changes from egg-liking to opening ,which decreases the commercial value of the fruit-body.What makes Volvariella volvacea fruit-body changed so much just in this short time? The cell morpha , internal hormones and the expression of special genes are studied in this paper.
1. Cell elongation reduces the rapid growth of fruit-body stipe in the elongation stage We observed cell morpha and measured cell size of different Volvariella volvacea fruit-body stipe and different parts of stipes in the egg and the elongation stages .At the same time we observed and measured the elongation region of fruit-body stipe by stipe labeled.The results showed : fruit-body stipes in egg stage grow and the quantity of the cells increases by cell splitting .The essential factor of the spread of fruit-body stipe has been the elongation of cell on the middle top of stipe since the elongation stage ,in the meanwhile the top cell splitting of stipe plays assistant role.
2.Significant relevancy between the change of endogenous hormone and fruit-body growth Determination of the lever change of endogenous hormone of fruit-body stipe at the different development stages shows:there are not significant changes whether growth hormone or senescent hormone.But in elongation stage .growth hormone OA3 and ZR increase dramatically and reach the peak, senescent hormone ABA declines sharply .In mature stage , although the content of GA3 and ZR are lower than those in elongation stage, which remain fairly high .The concentration of ABA reach the lowest lever.The ratio of GA3 and ZR has changed with the concentration change of GA3.the ratio is essential factor which affects the growth of stipe cells.But other growth factors that control sensecence are key effect during senescence from fruit-body to mature..
3.Cloning gene fragment expression specific in the opening stage
Total mRNA of fruit-body stipe is regarded as Tester and Driver respectively in egg stage and elongation stage to clone special expression gene fragment of fruit-body stipe in elongation stage by suppression subtraction hybridization.We got a gene fragment and sequence. We did not get the same sequence with our sequence by homology comparision in GENBANK. We took this fragment as template to synthesize primer and have two-dimensional amplification of genome DNA.The amplification with two primer gains target fragment ,at the same time , amplification with single primer get a fragment about 1-2KB.
4. Construction of expression vector of sense and antisense gene fragment
Target gene fragment in plasmid is insolated and cloned with enzyme digest.In the
2
meanwhile the target expression vector is dealt with end-filling and dephosphorylation. Cloned gene fragment inserts to the polycloning site of PCB vector by connection of blunt end, then transformed E.coli and selected positive clony at random.Plasmids were extracted and sequenced. We verified that we gained cloned vector with reciprocal sequences.
1．细胞伸长主导伸长期子实体菌柄快速伸长 显微观察草姑子实体菌柄细胞在不同发育时期以及菌柄不同部位在蛋形期和伸长期的细胞形态并测量了细胞大小变化，同时通过菌柄标记观察并测量了子实体菌柄的伸长区间。结果表明：在蛋形期草菇主要是通过细胞分裂使细胞数增加而使子实体长大，从伸长期开始，子实体菌柄伸长的主导因素是菌柄中上部细胞的伸长，同时菌柄顶部细胞的分裂起辅助作用。
2．内源激素水平变化与子实体生长显著相关 测量子实体菌柄在不同发育阶段的内源激素水平变化，结果显示：在蛋形期之前，无论促进生长类激素还是衰老类激素都没有明显的变化；但是在伸长期，促进生长类激素GA_(1/3)和ZRs急剧上升到顶峰，衰老类激素ABA则急剧下降；到成熟期，GA_(1/3)和ZRs的含量比伸长期时明显下降，但仍维持在较高的水平，ABA的浓度在成熟期时下降到最低水平。在不同发育时期中GA_(1/3)的浓度变化主导GA_(1/3)／ABA的比率变化，GA_(1/3)／ABA可能是主导菌柄细胞生长的重要因素，但子实体进入成熟期后的衰老过程可能有其它控制衰老的生理因子起关键作用。
3．克隆到开伞阶段特异表达基因片段 通过抑制性扣除杂交法，以蛋形期子实体菌柄的Total mRNA为驱赶子，伸长期子实体菌柄的Total mRNA为目的子，克隆子实体菌柄在伸长期特异表达的基因，结果得到一个基因片段，测序并登录GENBANK进行同源性比较，没有与此序列相同的基因。以该基因片段为模板合成引物并进行基因组DNA扩增，双引物扩增到目的片段，单引物扩增得到1～2KB左右的扩增片段。
4．正、反义基因片段表达载体的构建 酶切分离克隆质粒中的目的基因片段，同时对酶切的目的表达载体进行末端平滑及去磷酸化处理，通过平末端连接，将克隆的基因片段插入到PCB载体的多克隆位点中，转化大肠杆菌，随机挑选阳性菌落，提取质粒进行测序，证实获得分别具有正反序列的克隆载体。
The development of Volvariella volvacea fruit-body has six stages of the pinhead,the tinny button,the button,the egg,the elongation and the mature.The physiological characteristics of the fruit-body changes obviously in different stages, respiratiry intensity enhanced dramatically especially from the egg stage to the elongation stage.At the same time , the morpha of the fruit-body changes from egg-liking to opening ,which decreases the commercial value of the fruit-body.What makes Volvariella volvacea fruit-body changed so much just in this short time? The cell morpha , internal hormones and the expression of special genes are studied in this paper.
1. Cell elongation reduces the rapid growth of fruit-body stipe in the elongation stage We observed cell morpha and measured cell size of different Volvariella volvacea fruit-body stipe and different parts of stipes in the egg and the elongation stages .At the same time we observed and measured the elongation region of fruit-body stipe by stipe labeled.The results showed : fruit-body stipes in egg stage grow and the quantity of the cells increases by cell splitting .The essential factor of the spread of fruit-body stipe has been the elongation of cell on the middle top of stipe since the elongation stage ,in the meanwhile the top cell splitting of stipe plays assistant role.
2.Significant relevancy between the change of endogenous hormone and fruit-body growth Determination of the lever change of endogenous hormone of fruit-body stipe at the different development stages shows:there are not significant changes whether growth hormone or senescent hormone.But in elongation stage .growth hormone OA3 and ZR increase dramatically and reach the peak, senescent hormone ABA declines sharply .In mature stage , although the content of GA3 and ZR are lower than those in elongation stage, which remain fairly high .The concentration of ABA reach the lowest lever.The ratio of GA3 and ZR has changed with the concentration change of GA3.the ratio is essential factor which affects the growth of stipe cells.But other growth factors that control sensecence are key effect during senescence from fruit-body to mature..
3.Cloning gene fragment expression specific in the opening stage
Total mRNA of fruit-body stipe is regarded as Tester and Driver respectively in egg stage and elongation stage to clone special expression gene fragment of fruit-body stipe in elongation stage by suppression subtraction hybridization.We got a gene fragment and sequence. We did not get the same sequence with our sequence by homology comparision in GENBANK. We took this fragment as template to synthesize primer and have two-dimensional amplification of genome DNA.The amplification with two primer gains target fragment ,at the same time , amplification with single primer get a fragment about 1-2KB.
4. Construction of expression vector of sense and antisense gene fragment
Target gene fragment in plasmid is insolated and cloned with enzyme digest.In the
2
meanwhile the target expression vector is dealt with end-filling and dephosphorylation. Cloned gene fragment inserts to the polycloning site of PCB vector by connection of blunt end, then transformed E.coli and selected positive clony at random.Plasmids were extracted and sequenced. We verified that we gained cloned vector with reciprocal sequences.
引文
[1] Chang.S.T.and C.K.Yau, Volvariella volavacea and its life history Ameri.J.Bot.1971, 58: 552～561
[2] 张树庭，1975《草菇》：33～34
[3] 郭家选，赵永厚等，几种食用菌菌丝呼吸生理的研究，中国生态农业学报，2002，10(3)：74～75
[4] 邓茂常，草菇栽培技术，资源开发与市场，1994，10(1)：31
[5] 叶蕙，陈建勋，余让才等，60Coγ辐照对草菇保鲜及其生理机制的研究，核农学报2000，10(6)：1～6
[6] 陈金印，陈明，果实后熟衰老与植物激素的关系研究进展，江西农业大学学报，2003，25(4)：537～543
[7] 辜诗玲，浅谈草菇栽培，中国食用菌，1998，17(6)：35～36
[8] 谢宝贵，江玉姬，草菇厚垣孢子研究，食用菌学报，1996，3(4)：25～29
[9] 石维民，草菇栽培中的问题与分析，山西农业科学，1990(9)：24～25
[10] 周竹清，草菇栽培七大问题的预防对策，技术开发与引进，1993(3)：25～26
[11] 韦文添，草菇栽培中生长异常的原因及预防措施，蔬菜，2003(12)：20～21
[12] 张闽春，李连伍，低温季节草菇栽培关键技术，北京农业，2001，(11)：22～22
[13] 张财先，草菇栽培过程中鬼伞发生的原因及防治技术，中国食用菌，2000，19(1)：33～34
[14] 杨佩玉，陈上海，室内草菇栽培工艺的研究，食用菌，1991，13(1)：32～33
[15] 齐辉，日光温室越夏草姑栽培，农村实用工程技术，2002，(9)：11～11
[16] 温海祥，肖洪东，草菇品种间酶活性差异的研究，中国食用菌，1999，18(5)：23～24
[17] 万鲁长，兰传进，北方地区麦桔复合料有效积温与草菇生物量关系研究初报，山东农业科学，1999(1)：12～14
[18] 王春晖，张志光，外源DNA片段导入草菇原生质体的研究，食用菌学报，1999，6(4)：1～6
[19] 邹伯琼，曹裕汉，草菇单孢子选育种研究，广东农业科学，2001(4)：23～24
[20] 曹裕汉，草菇有性分离选育性研究，食用菌，2003，25(2)：12～13
[21] Chang S T, Hayes W A, et.al, The biology and cultivation of edible mushroom, New York: Academia Press, 1978:573～605
[22] Chang S T, Buswell J A, Chiu S W, et al, Mushroom biology and mushroom products,
Hong Kong: The Chinese University Press, 1993:71～83
[23] 董华强等，室温臭氧保鲜草菇技术，食用菌，1998，(6)：35～36
[24] 董华强等，室温臭氧保鲜草菇研究，中国食用菌，1999，(2)：40～41
[25] 王富民，宫秀荣，高君辉等，草菇保鲜技术及有关生理变化，上海农业学报，1992，8(3)：60～68
[26] 叶蕙，陈建勋，γ辐照对草菇保鲜及其生理机制的研究，核农学报，2000，14(1)：24-28
[27] 郑海歌，陈明杰，顾向红，草菇发育过程中某些衰老因子的变化，上海农业学报，1991，7(2)：27～31
[28] 戴志刚，陈明杰，潘迎捷，草菇低温诱导蛋白分离纯化及其部分性质的测定，菌物系统，2003，22(4)：579～583
[29] 陈明杰，谭琦等，mRNA差别显示技术分离草菇低温诱导基因，菌物系统，2001，20(3)：342～346
[30] 郭丽琼，林俊芳等，THP基因的重新克隆及草菇表达载体的构建，微生物学报，2002，42(3)：375～379
[31] Chen S, Ge W, Buswell JA., Molecular cloning of a new laccase from the edible straw mushroom Volvariella volvacea: possible involvement in fruit body development, FEMS Microbiol Lett. 2004 Jan 30; 230(2): 171-6
[32] Chen S, Ge W, Buswell JA., Biochemical and molecular characterization of a laccase from the edible straw mushroom, Volvariella volvacea, Eur J Biochem. 2004 Jan; 271(2): 318-28.
[33] Jia J, Dyer PS, Buswell JA, Peberdy JF., Cloning of the cbhl and cbhll genes involved in cellulose utilisation by the straw mushroom Volvariella volvacea, Mol Gen Genet. 1999 Jul; 261(6): 985-93.
[34] Yao QZ, Yu MM, Ooi LS .et. Al, Isolation and Characterization of a Type 1 Ribosome-Inactivating Protein from Fruiting Bodies of the Edible Mushroom (Volvariella volvacea), J Agric Food Chem. 1998 Feb 16; 46(2): 788-792
[35] Liua W, Ho JC, Ng T, Suppression of cell cycle progression by a fungal lectin: activation of cyclin-dependent kinase inhibitors, Biochem Pharmacol. 2001 Jan 1, 61 (1): 33～37
[36] She QB, Ng TB, Liu WK, A novel lectin with potent immunomodulatory activity isolated from both fruiting bodies and cultured mycelia of the edible mushroom Volvariella volvacea, Biochem Biophys Res Commun. 1998 Jun 9, 47(1): 106～111
[37] Hsu HC, Hsu CI, Lin RH.et. al, Fip-vvo, a new fungal immunomodulatory protein isolated
from Volvariella volvacea, Biocbem J. 1997 Apr 15, 323 (Pt 2): 557-65
[38] Sone Y, Shibata S, Kishida E, Alteration of beta-D-glucan from edible mushroom after injection into mouse peritoneal cavity, J Nutr Sci Vitaminol (Tokyo). 1994 Aug; 40(4): 335-41
[39] Kishida E, Kinoshita C, Sone Y, Misaki A, Structures and antitumor activities of polysaccharides isolated from mycelium of Volvariella voivacea, Biosci Biotechnol Biochem. 1992 Aug, 56(8): 1308-1309
[40] 王三根，王西瑶，1997《植物生理学》：202～241
[41] 孙振令，张红雨，芸苔素内酯的研究进展及其在农业生产中的应用，淄博学院学报(自然科学与工程版)，2001，3(2)：81～84
[42] 王玉琴，周维善，油菜甾醇内酯类化合物生物活性与构效关系的研究，科学通报，1994，39(7)：649～652
[43] 郑玉生，张秋明等，多胺类代谢与园艺作物生长发育关系研究进展，亚热带植物通讯，1996，25(1)：43～50
[44] 李劲，萧浪涛等，植物内源茉莉酸类生长物质研究进展，湖南农业大学学报(自然科学版)，2002，28(1)：78～84
[45] 谷丽萍，周阮宝，茉莉酸类在植物体内作用研究进展，植物学通报，1997，14(1)：31-35
[46] 岳寿松，于振文，余松烈，小麦旗叶衰老过程中氧自由基与激素含量的变化(简报)，植物生理学通讯，1996，32(5)：349～351
[47] 张志华，王文江，核桃果实成熟过程中呼吸速率与内源激素的变化，园艺学报，2000，27(3)：167～170
[48] 朱中华，段留生，冯雪梅等，内源激素对小麦叶片衰老调控的系统分析，作物学报，1998，24(2)：176～181
[49] 吴邦平，激素对平菇菌丝生长及产量的影响，食用菌，1989，11(6)：18～18
[50] 刘喜文，王金胜激素在猴头菌丝体碳源利用过程中的作用，山西农业大学学报(自然科学版)，1994，14(3)：269～271
[51] 齐国辉，郗荣庭，VA菌根菌对苹果组培苗内源激素含量的影响，河北农业大学学报，1997，20(4)：51～54
[52] 王有智，黄亦存，四种外生菌根真菌产生植物激素的研究，微生物学通报，1997，24(2)：72～74
[53] 檀根甲，承河元等，几种抗菌素和激素类物质对稻白叶枯病菌生长的影响，安徽农
业科学，1997，25(3)：260～271
[54] LIU Run Jin, LI Min et al., EFFECTS OF AM FUNGI ON ENDOGENOUS HORMONES IN CORN AND COTTON PLANTS, Mycosustema, 200, 19(1): 91～96
[55] 吴克刚，柴向华，杨连生，植物激素对破囊壶菌生长与产DHA的影响，微生物学报，2003，43(1)：111-115
[56] Liang P, Pardee AB., Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction, Science. 1992 Aug 14; 257(5072): 967～71
[57] Welsh J, Chada K, Dalal SS, Cheng R, Ralph D, McClelland M., Arbitrarily primed PCR fingerprinting of RNA, Nucleic Acids Res. 1992 Oct 11: 20(19): 4965～70.
[58] Lisitsyn N, Lisitsyn N, Wigler M., Cloning the differences between two complex genomes, Science. 1993 Feb 12; 259(5097): 946-51
[59] 孙春晓，于常海，基因克隆的常用方法介绍，生理科学进展，2000，31(1)：35～42
[60] Diatchenko L,Lau Y C,Campbell A P, et al.Proc Natl Acad Sci USA 1996,93:6025～6030
[61] Van Stein OD,et al.Nucleic Acids research, 1997,25(13):2598～2602.
[62] Lee JH, Miele ME, Hicks DJ et al.J Natl Cancer Inst, 1996, 88(23)1731-1737
[63] Garkavtsev I Kazarov A, Gudkov Aet al.Nat Genet, 1996, 14(4): 415～420
[64] Liu XL, Wazer DE, Watannabe K et al.Cancer Res, 1996, 56(14)3371～3379
[65] Shimada T, Matsumoto M, Tatsumi Y et al, FEBS lett, 1998, 425(3): 490～194
[66] Nemeth E.Bole-Feysot C, Tashima LS.J Mol Endocrinol, 1998.20(1): 151～156
[67] Schiniel C, Pardai E, Sallusto F et al.J Exp Med, 1998, 188(3): 451～463
[68] Guan LS, Rauchman M, Wang ZY et al.J Biol Chem, 1998, 273(42): 27047～27050
[69] Thompson DA, Weigel RJ, Biochem Bionphys Res Commun, 1998, 251(1): 111～116
[70] XIA Ming, WANG Xiao-Bing, LI Hai-Bo, WU Ping, Identification of the Rice Vacuolar ATPase B Subunit Gene and Its Expression Pattern Analusis Under Phosphorus Deficiency, Acta Botanica, 2002, 44(5): 573～578
[71] 张咏，崔春萍，鱼咏涛等，长期培养后小鼠造血基质细胞差异表达基因的筛选，中国实验血液学杂志，2002，10(3)：177-182
[72] 张必成，周洁，朱诗国等，鼻咽癌上皮细胞株HNE1差异表达基因的分离与鉴定，中国生物化学与分子生物学报，2003，19(6)：743～750
[73] Shim KS, Cho SK, Jeung JU, Identification of fungal (Magnaporthe grisea) stress-induced genes in wild rice ( Oryza minuta),: Plant Cell Rep. 2004 Mar; 22(8): 599-607. Epub 2003 Nov 27
[74] Xie D, Jauch A, Miller CW, Bantam CR, Koeffler HP, Discovery of over-expressed genes and genetic alterations in breast cancer cells using a combination of suppression subtractive hybridization, multiplex FISH and comparative genomic hybridization, Int J Oncol. 2002 Sep: 21(3): 499-507.
[75] Rimner A, Wischhusen J, Naumann U, Gleichmann M, Steinbach JP, Weller M, Identification by suppression subtractive hybridization of p21 as a radio-inducible gene in human glioma cells, Anticancer Res. 2001 Sep-Oct; 21(5): 3505-8
[76] Den Hollander AI, et al.Genomics, 1999, 58(3): 240～249
[77] Nickel Al, et al.Int Arch Allergy Immumol, 1999, 118(24): 282～284
[78] Sun L, et al.J Biol Chem., 1999, 274(24): 17334～17341
[79] George P, Yang, et al, Nucleic Acids Research, 1999, 27(6): 1517～1523
[80] Wu GS, Saftig P, Peters C et al. Oncogene, 1998, 16(17)2177～2183
[81] Bovre K, Szybalski W, Patterns of convergent and overlapping transcription within the b2 region ofcoliphage lambda.Virology. 1969 Aug; 38(4): 614-26
[82] Itoh T, Tomizawa J. Formation of an RNA primer for initiation of replication of ColE1 DNA by ribonuclease H.Proc Natl Acad Sci U S A. 1980 May; 77(5): 2450-4
[83] Simons RW, Kleckner N, Translational control of IS10 transposition.Cell. 1983 Sep; 34(2): 683～91
[84] 王敏，卵巢恶性肿瘤的反义基因治疗研究进展，国外医学：1998，25(5)：285～289
[85] Cirino NM, Li G, Xiao W, etal., Targeting RNA decay with 2', 5'-oligoadenylate antisense in tespiratory syncytial virua-infected cells[J] Proc, Natl.Acad.Sci., 1997, 94(5): 1937～1942
[86] 朱小石，李缠生，呼吸道病毒基因反义技术的研究进展，国外医学病毒学分册，2001，8(1)：19～22
[87] Helm CW et al, Gynecol Oncol, 1993, 49:339-343
[88] Postel EH, Flint SJ, Kessler DJ, et al.Evidence that a triplex-forming oligodeoxyribo-nucleotide binds to the c-myc mRNA levels[J] Proc Natl Acad Sci USA, 1991, 88(18): 8227～8231
[89] Gu ZP, Wang YJ, Li JG, et al.VEGG165 antisense RNA suppression oncogenic properties of human esophageal squamous cell carcinoma[J], World J Gastroenterol, 2002, 8(1): 44
[90] Jae J Song, Heuiran Lee, Eunhee Kim, et al.Transduction effect of antisense K-ras on malignant phenotypesin gastric cancer cells[J] , Cancer Letters, 2000, 157(1): 1.
[91] Ren J, Dong L, Xu CB, et al., The role of KDR in the interaction between human gastric carcinoma cell and vascular endothelial cell[J], World J Gastroenterol, 2002, 8(4): 596.
[92] Uto H, Ido A, Moriuchi A, et al.Transduction of antisense cyclin D1 using two-step gene transfer inhibits the growth of rat hepatoma cells[J], Cancer Res, 2001, 15; 61(12)4779
[93] 司遂海，杨建民，罗元辉等，KAⅡ正反义基因对MHCC97-H肝癌细胞KAⅡ蛋白表达的影响，世界华人消化杂志，2003，11(9)：1341～1344
[94] Bedi A, Zehnbauer BA, Barber JP, Sharkis SJ, Jones RJ. Inhibition of apoptosis by BCR-ABL in chronic myeloid leukemia.Blood. 1994 Apr 15; 83(8): 2038～44.
[95] 张凤春，林玉梅，反义基因逆转肿瘤细胞多药耐药性诱导细胞凋亡的研究，暨南大学学报(自然科学与医学版)，1998，19(6)：85～86
[96] 张宏，肖文华等，人DNA MTase反义基因转染对诱导肝癌SMMC—7721细胞凋亡的影响，第三军医大学学报，2002，24(11)：1270～1272
[97] 龚海霞，郭锡熔等，神经肽Y—Y5受体反义基因治疗诱导大鼠脂肪细胞凋亡研究，Journal of Nanjing Medical University(南京医科大学学报：英文版)，2003，17(1)：1～9
[98] van der Krol AR, Mol JN, Stuitje AR, Antisense genes in plants: an overview, Gene. 1988 Dec 10; 72(1-2): 45-50.
[99] Smith CJ, Watson CF, Morris PC et al, Inheritance and effect on ripening of antisense polygalacturonase genes in transgenic tomatoes, Plant Mol Biol. 1990 Mar; 14(3): 369-79.
[100] Smith CJ, Watson CF, Bird CR, Ray J, Schuch W, Grierson D., Expression of a truncated tomato polygalacturonase gene inhibits expression of the endogenous gene in transgenic plants.Mol Gen Genet. 1990 Dec; 224(3): 477-81
[101] P A Powell, D M Stark, P R Sanders, and R N Beachy , Protection against tobacco mosaic virus in transgenic plants that express tobacco mosaic virus antisense RNA, Proc Natl Aead Sci U S A. 1989 September; 86 (18): 6949-6952
[102] Soo-Hwan Kim, David Arnold, Alan Lloyd, and Stanley J. Roux, Antisense Expression of an Arabidopsis Ran Binding Protein Renders Transgenic Roots Hypersensitive to Auxin and Alters Auxin-Induced Root Growth and Development by Arresting Mitotic Progress
[103] 党伟，吕万革，蕃茄多聚半乳糖醛酸酶的cDNA克隆及反义基因转化植物，Acta Botanica Sinica(植物学报：英文版)，1994(A 11)：171～177
[104] 马庆虎，宋艳茹，多聚半乳糖醛酸酶反义基因在转基因番茄中的表达，生物多样性，1995，3(1)：21～25
[105] 叶志彪，李汉霞，反义ACC氧化酶基因的导入对番茄乙烯生成的抑制作用，华中农业大学学报，1996，15(4)：305～309
[106] 王春霞，简志英，ACC合成酶基因及其反义基因对西瓜的遗传转化，Acta Botanica Siniea(植物学报：英文版)，1997，39(5)：445～450
[107] 陈锦清，郎春秀，反义PEP基因调控油菜籽粒蛋白质／油脂含量比率的研究，农业生物技术学报，1999，7(4)：316～320
[108] 郊娥香，杨志行等，水稻bicoid反义基因转基因植株胚胎发育的形态变化，植物学通报，2001，18(6)：707～713
[109] Fire A, Xu S, Montgomery MK, Kostas SA, et al, Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans., Nature, 1998, 391 (6669): 806～811
[110] Guo S, Kemphues KJ., par-1, a gene required for establishing polarity in C. elegans embryos, encodes a putative Ser/Thr kinase that is asymmetrically distributed, Cell. 1995 May 19; 81(4): 611-20. Cell. 1995 May 19; 81(4): 611～20
[111] 马鹏鹏，薛社普，韩代书，RNA干扰技术的原理与应用，中国组织化学与细胞化学杂志，2003，12(2)：201～207
[112] Brantl S., Antisense-RNA regulation and RNA interference, Biochim Biophys Acta. 2002 May3; 1575(1-3): 15～25
[113] Colaiacovo MP, Stanfield GM, Reddy KC, Reinke V, Kim SK, Villeneuve AM., A targeted RNAi screen for genes involved in chromosome morphogenesis and nuclear organization in the Caenorhabditis elegans germline, Genetics. 2002 Sep; 162(1): 113～28
[114] Yu JY, DeRuiter SL, Turner DL., RNA interference by expression of short-interfering RNAs and hairpin RNAs in mammalian cells, Proc Natl Acad Sci U S A. 2002 Apr 30; 99(9): 6047～6052.
[115] Czaudema F, Fechtner M, Aygun H, Arnold W, Klippel A, Giese K, Kaufmann J., Functional studies of the PI(3)-kinase signalling pathway employing synthetic and expressed siRNA.Nucleic Acids Res. 2003 Jan 15; 31(2): 670～682.
[116] 朱晓燕，刘宇健等，利用载体介导的RNA干扰技术抑制小鼠腹腔巨噬细胞RAW264．7内糖皮质激素受体表达及功能的研究，中国病理生理杂志，2003，19(11)：1541～1542
[117] Yu Shu-wen, Tang Zhang-cheng: Plant Physiology and Molecular Biology (2ed). Science Press. 1998.
[118] Forde, J., Malpica, J.M., The nucleotide sequence of a HMW glutenin subunit gene located on chromosome 1A of wheat (Triticum aestivum L.), Nucleic Acids Res. 13 (19), 6817-6832 (1985)
[119] Gu, Y.Q., Coleman-Derr, D.A., Rapid Genome Evolution Revealed By Comparative Analysis Of Orthologous Regions From Four Triticeae Genomes
[120] Chapman, J., Direct Submission(GeneBank), Apr 11, 2003
[121] Bhairi S M, Staples R C. 1992. Transient expression of the β-Glucuronidase Gene introduced into Uromyces appendicμlatus uredospores by particle bombardment. Phytopathology. 82(9):986～989
[122] Chen X, Stone M, Schlagnhaufer C, Romaine C P. 2000. A fruiting body tissue method for efficient Agrobacterium mediated transformation of Agaricus bisporus. Applied and Environmental Microbiology. 66(10):4510～4513
[123] Smith CJ, Watson CF, Morris PC, et, al, Inheritance and effect on ripening of antisense polygalacturonase genes in transgenic tomatoes, Plant Mol Biol. 1990 Mar: 14(3) :369-79
[124] Tentler S, Palas J, Enderlin C, et al. 1997. Inhibition of Neurospora crassa growth by a glucan synthaser-1 antisense construct. Current Microbiology. 34:303～308
[125] (美)J．萨姆布鲁克等，《分子克隆》第三版，2002：96～99，103～105，26～35
[2] 张树庭，1975《草菇》：33～34
[3] 郭家选，赵永厚等，几种食用菌菌丝呼吸生理的研究，中国生态农业学报，2002，10(3)：74～75
[4] 邓茂常，草菇栽培技术，资源开发与市场，1994，10(1)：31
[5] 叶蕙，陈建勋，余让才等，60Coγ辐照对草菇保鲜及其生理机制的研究，核农学报2000，10(6)：1～6
[6] 陈金印，陈明，果实后熟衰老与植物激素的关系研究进展，江西农业大学学报，2003，25(4)：537～543
[7] 辜诗玲，浅谈草菇栽培，中国食用菌，1998，17(6)：35～36
[8] 谢宝贵，江玉姬，草菇厚垣孢子研究，食用菌学报，1996，3(4)：25～29
[9] 石维民，草菇栽培中的问题与分析，山西农业科学，1990(9)：24～25
[10] 周竹清，草菇栽培七大问题的预防对策，技术开发与引进，1993(3)：25～26
[11] 韦文添，草菇栽培中生长异常的原因及预防措施，蔬菜，2003(12)：20～21
[12] 张闽春，李连伍，低温季节草菇栽培关键技术，北京农业，2001，(11)：22～22
[13] 张财先，草菇栽培过程中鬼伞发生的原因及防治技术，中国食用菌，2000，19(1)：33～34
[14] 杨佩玉，陈上海，室内草菇栽培工艺的研究，食用菌，1991，13(1)：32～33
[15] 齐辉，日光温室越夏草姑栽培，农村实用工程技术，2002，(9)：11～11
[16] 温海祥，肖洪东，草菇品种间酶活性差异的研究，中国食用菌，1999，18(5)：23～24
[17] 万鲁长，兰传进，北方地区麦桔复合料有效积温与草菇生物量关系研究初报，山东农业科学，1999(1)：12～14
[18] 王春晖，张志光，外源DNA片段导入草菇原生质体的研究，食用菌学报，1999，6(4)：1～6
[19] 邹伯琼，曹裕汉，草菇单孢子选育种研究，广东农业科学，2001(4)：23～24
[20] 曹裕汉，草菇有性分离选育性研究，食用菌，2003，25(2)：12～13
[21] Chang S T, Hayes W A, et.al, The biology and cultivation of edible mushroom, New York: Academia Press, 1978:573～605
[22] Chang S T, Buswell J A, Chiu S W, et al, Mushroom biology and mushroom products,
Hong Kong: The Chinese University Press, 1993:71～83
[23] 董华强等，室温臭氧保鲜草菇技术，食用菌，1998，(6)：35～36
[24] 董华强等，室温臭氧保鲜草菇研究，中国食用菌，1999，(2)：40～41
[25] 王富民，宫秀荣，高君辉等，草菇保鲜技术及有关生理变化，上海农业学报，1992，8(3)：60～68
[26] 叶蕙，陈建勋，γ辐照对草菇保鲜及其生理机制的研究，核农学报，2000，14(1)：24-28
[27] 郑海歌，陈明杰，顾向红，草菇发育过程中某些衰老因子的变化，上海农业学报，1991，7(2)：27～31
[28] 戴志刚，陈明杰，潘迎捷，草菇低温诱导蛋白分离纯化及其部分性质的测定，菌物系统，2003，22(4)：579～583
[29] 陈明杰，谭琦等，mRNA差别显示技术分离草菇低温诱导基因，菌物系统，2001，20(3)：342～346
[30] 郭丽琼，林俊芳等，THP基因的重新克隆及草菇表达载体的构建，微生物学报，2002，42(3)：375～379
[31] Chen S, Ge W, Buswell JA., Molecular cloning of a new laccase from the edible straw mushroom Volvariella volvacea: possible involvement in fruit body development, FEMS Microbiol Lett. 2004 Jan 30; 230(2): 171-6
[32] Chen S, Ge W, Buswell JA., Biochemical and molecular characterization of a laccase from the edible straw mushroom, Volvariella volvacea, Eur J Biochem. 2004 Jan; 271(2): 318-28.
[33] Jia J, Dyer PS, Buswell JA, Peberdy JF., Cloning of the cbhl and cbhll genes involved in cellulose utilisation by the straw mushroom Volvariella volvacea, Mol Gen Genet. 1999 Jul; 261(6): 985-93.
[34] Yao QZ, Yu MM, Ooi LS .et. Al, Isolation and Characterization of a Type 1 Ribosome-Inactivating Protein from Fruiting Bodies of the Edible Mushroom (Volvariella volvacea), J Agric Food Chem. 1998 Feb 16; 46(2): 788-792
[35] Liua W, Ho JC, Ng T, Suppression of cell cycle progression by a fungal lectin: activation of cyclin-dependent kinase inhibitors, Biochem Pharmacol. 2001 Jan 1, 61 (1): 33～37
[36] She QB, Ng TB, Liu WK, A novel lectin with potent immunomodulatory activity isolated from both fruiting bodies and cultured mycelia of the edible mushroom Volvariella volvacea, Biochem Biophys Res Commun. 1998 Jun 9, 47(1): 106～111
[37] Hsu HC, Hsu CI, Lin RH.et. al, Fip-vvo, a new fungal immunomodulatory protein isolated
from Volvariella volvacea, Biocbem J. 1997 Apr 15, 323 (Pt 2): 557-65
[38] Sone Y, Shibata S, Kishida E, Alteration of beta-D-glucan from edible mushroom after injection into mouse peritoneal cavity, J Nutr Sci Vitaminol (Tokyo). 1994 Aug; 40(4): 335-41
[39] Kishida E, Kinoshita C, Sone Y, Misaki A, Structures and antitumor activities of polysaccharides isolated from mycelium of Volvariella voivacea, Biosci Biotechnol Biochem. 1992 Aug, 56(8): 1308-1309
[40] 王三根，王西瑶，1997《植物生理学》：202～241
[41] 孙振令，张红雨，芸苔素内酯的研究进展及其在农业生产中的应用，淄博学院学报(自然科学与工程版)，2001，3(2)：81～84
[42] 王玉琴，周维善，油菜甾醇内酯类化合物生物活性与构效关系的研究，科学通报，1994，39(7)：649～652
[43] 郑玉生，张秋明等，多胺类代谢与园艺作物生长发育关系研究进展，亚热带植物通讯，1996，25(1)：43～50
[44] 李劲，萧浪涛等，植物内源茉莉酸类生长物质研究进展，湖南农业大学学报(自然科学版)，2002，28(1)：78～84
[45] 谷丽萍，周阮宝，茉莉酸类在植物体内作用研究进展，植物学通报，1997，14(1)：31-35
[46] 岳寿松，于振文，余松烈，小麦旗叶衰老过程中氧自由基与激素含量的变化(简报)，植物生理学通讯，1996，32(5)：349～351
[47] 张志华，王文江，核桃果实成熟过程中呼吸速率与内源激素的变化，园艺学报，2000，27(3)：167～170
[48] 朱中华，段留生，冯雪梅等，内源激素对小麦叶片衰老调控的系统分析，作物学报，1998，24(2)：176～181
[49] 吴邦平，激素对平菇菌丝生长及产量的影响，食用菌，1989，11(6)：18～18
[50] 刘喜文，王金胜激素在猴头菌丝体碳源利用过程中的作用，山西农业大学学报(自然科学版)，1994，14(3)：269～271
[51] 齐国辉，郗荣庭，VA菌根菌对苹果组培苗内源激素含量的影响，河北农业大学学报，1997，20(4)：51～54
[52] 王有智，黄亦存，四种外生菌根真菌产生植物激素的研究，微生物学通报，1997，24(2)：72～74
[53] 檀根甲，承河元等，几种抗菌素和激素类物质对稻白叶枯病菌生长的影响，安徽农
业科学，1997，25(3)：260～271
[54] LIU Run Jin, LI Min et al., EFFECTS OF AM FUNGI ON ENDOGENOUS HORMONES IN CORN AND COTTON PLANTS, Mycosustema, 200, 19(1): 91～96
[55] 吴克刚，柴向华，杨连生，植物激素对破囊壶菌生长与产DHA的影响，微生物学报，2003，43(1)：111-115
[56] Liang P, Pardee AB., Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction, Science. 1992 Aug 14; 257(5072): 967～71
[57] Welsh J, Chada K, Dalal SS, Cheng R, Ralph D, McClelland M., Arbitrarily primed PCR fingerprinting of RNA, Nucleic Acids Res. 1992 Oct 11: 20(19): 4965～70.
[58] Lisitsyn N, Lisitsyn N, Wigler M., Cloning the differences between two complex genomes, Science. 1993 Feb 12; 259(5097): 946-51
[59] 孙春晓，于常海，基因克隆的常用方法介绍，生理科学进展，2000，31(1)：35～42
[60] Diatchenko L,Lau Y C,Campbell A P, et al.Proc Natl Acad Sci USA 1996,93:6025～6030
[61] Van Stein OD,et al.Nucleic Acids research, 1997,25(13):2598～2602.
[62] Lee JH, Miele ME, Hicks DJ et al.J Natl Cancer Inst, 1996, 88(23)1731-1737
[63] Garkavtsev I Kazarov A, Gudkov Aet al.Nat Genet, 1996, 14(4): 415～420
[64] Liu XL, Wazer DE, Watannabe K et al.Cancer Res, 1996, 56(14)3371～3379
[65] Shimada T, Matsumoto M, Tatsumi Y et al, FEBS lett, 1998, 425(3): 490～194
[66] Nemeth E.Bole-Feysot C, Tashima LS.J Mol Endocrinol, 1998.20(1): 151～156
[67] Schiniel C, Pardai E, Sallusto F et al.J Exp Med, 1998, 188(3): 451～463
[68] Guan LS, Rauchman M, Wang ZY et al.J Biol Chem, 1998, 273(42): 27047～27050
[69] Thompson DA, Weigel RJ, Biochem Bionphys Res Commun, 1998, 251(1): 111～116
[70] XIA Ming, WANG Xiao-Bing, LI Hai-Bo, WU Ping, Identification of the Rice Vacuolar ATPase B Subunit Gene and Its Expression Pattern Analusis Under Phosphorus Deficiency, Acta Botanica, 2002, 44(5): 573～578
[71] 张咏，崔春萍，鱼咏涛等，长期培养后小鼠造血基质细胞差异表达基因的筛选，中国实验血液学杂志，2002，10(3)：177-182
[72] 张必成，周洁，朱诗国等，鼻咽癌上皮细胞株HNE1差异表达基因的分离与鉴定，中国生物化学与分子生物学报，2003，19(6)：743～750
[73] Shim KS, Cho SK, Jeung JU, Identification of fungal (Magnaporthe grisea) stress-induced genes in wild rice ( Oryza minuta),: Plant Cell Rep. 2004 Mar; 22(8): 599-607. Epub 2003 Nov 27
[74] Xie D, Jauch A, Miller CW, Bantam CR, Koeffler HP, Discovery of over-expressed genes and genetic alterations in breast cancer cells using a combination of suppression subtractive hybridization, multiplex FISH and comparative genomic hybridization, Int J Oncol. 2002 Sep: 21(3): 499-507.
[75] Rimner A, Wischhusen J, Naumann U, Gleichmann M, Steinbach JP, Weller M, Identification by suppression subtractive hybridization of p21 as a radio-inducible gene in human glioma cells, Anticancer Res. 2001 Sep-Oct; 21(5): 3505-8
[76] Den Hollander AI, et al.Genomics, 1999, 58(3): 240～249
[77] Nickel Al, et al.Int Arch Allergy Immumol, 1999, 118(24): 282～284
[78] Sun L, et al.J Biol Chem., 1999, 274(24): 17334～17341
[79] George P, Yang, et al, Nucleic Acids Research, 1999, 27(6): 1517～1523
[80] Wu GS, Saftig P, Peters C et al. Oncogene, 1998, 16(17)2177～2183
[81] Bovre K, Szybalski W, Patterns of convergent and overlapping transcription within the b2 region ofcoliphage lambda.Virology. 1969 Aug; 38(4): 614-26
[82] Itoh T, Tomizawa J. Formation of an RNA primer for initiation of replication of ColE1 DNA by ribonuclease H.Proc Natl Acad Sci U S A. 1980 May; 77(5): 2450-4
[83] Simons RW, Kleckner N, Translational control of IS10 transposition.Cell. 1983 Sep; 34(2): 683～91
[84] 王敏，卵巢恶性肿瘤的反义基因治疗研究进展，国外医学：1998，25(5)：285～289
[85] Cirino NM, Li G, Xiao W, etal., Targeting RNA decay with 2', 5'-oligoadenylate antisense in tespiratory syncytial virua-infected cells[J] Proc, Natl.Acad.Sci., 1997, 94(5): 1937～1942
[86] 朱小石，李缠生，呼吸道病毒基因反义技术的研究进展，国外医学病毒学分册，2001，8(1)：19～22
[87] Helm CW et al, Gynecol Oncol, 1993, 49:339-343
[88] Postel EH, Flint SJ, Kessler DJ, et al.Evidence that a triplex-forming oligodeoxyribo-nucleotide binds to the c-myc mRNA levels[J] Proc Natl Acad Sci USA, 1991, 88(18): 8227～8231
[89] Gu ZP, Wang YJ, Li JG, et al.VEGG165 antisense RNA suppression oncogenic properties of human esophageal squamous cell carcinoma[J], World J Gastroenterol, 2002, 8(1): 44
[90] Jae J Song, Heuiran Lee, Eunhee Kim, et al.Transduction effect of antisense K-ras on malignant phenotypesin gastric cancer cells[J] , Cancer Letters, 2000, 157(1): 1.
[91] Ren J, Dong L, Xu CB, et al., The role of KDR in the interaction between human gastric carcinoma cell and vascular endothelial cell[J], World J Gastroenterol, 2002, 8(4): 596.
[92] Uto H, Ido A, Moriuchi A, et al.Transduction of antisense cyclin D1 using two-step gene transfer inhibits the growth of rat hepatoma cells[J], Cancer Res, 2001, 15; 61(12)4779
[93] 司遂海，杨建民，罗元辉等，KAⅡ正反义基因对MHCC97-H肝癌细胞KAⅡ蛋白表达的影响，世界华人消化杂志，2003，11(9)：1341～1344
[94] Bedi A, Zehnbauer BA, Barber JP, Sharkis SJ, Jones RJ. Inhibition of apoptosis by BCR-ABL in chronic myeloid leukemia.Blood. 1994 Apr 15; 83(8): 2038～44.
[95] 张凤春，林玉梅，反义基因逆转肿瘤细胞多药耐药性诱导细胞凋亡的研究，暨南大学学报(自然科学与医学版)，1998，19(6)：85～86
[96] 张宏，肖文华等，人DNA MTase反义基因转染对诱导肝癌SMMC—7721细胞凋亡的影响，第三军医大学学报，2002，24(11)：1270～1272
[97] 龚海霞，郭锡熔等，神经肽Y—Y5受体反义基因治疗诱导大鼠脂肪细胞凋亡研究，Journal of Nanjing Medical University(南京医科大学学报：英文版)，2003，17(1)：1～9
[98] van der Krol AR, Mol JN, Stuitje AR, Antisense genes in plants: an overview, Gene. 1988 Dec 10; 72(1-2): 45-50.
[99] Smith CJ, Watson CF, Morris PC et al, Inheritance and effect on ripening of antisense polygalacturonase genes in transgenic tomatoes, Plant Mol Biol. 1990 Mar; 14(3): 369-79.
[100] Smith CJ, Watson CF, Bird CR, Ray J, Schuch W, Grierson D., Expression of a truncated tomato polygalacturonase gene inhibits expression of the endogenous gene in transgenic plants.Mol Gen Genet. 1990 Dec; 224(3): 477-81
[101] P A Powell, D M Stark, P R Sanders, and R N Beachy , Protection against tobacco mosaic virus in transgenic plants that express tobacco mosaic virus antisense RNA, Proc Natl Aead Sci U S A. 1989 September; 86 (18): 6949-6952
[102] Soo-Hwan Kim, David Arnold, Alan Lloyd, and Stanley J. Roux, Antisense Expression of an Arabidopsis Ran Binding Protein Renders Transgenic Roots Hypersensitive to Auxin and Alters Auxin-Induced Root Growth and Development by Arresting Mitotic Progress
[103] 党伟，吕万革，蕃茄多聚半乳糖醛酸酶的cDNA克隆及反义基因转化植物，Acta Botanica Sinica(植物学报：英文版)，1994(A 11)：171～177
[104] 马庆虎，宋艳茹，多聚半乳糖醛酸酶反义基因在转基因番茄中的表达，生物多样性，1995，3(1)：21～25
[105] 叶志彪，李汉霞，反义ACC氧化酶基因的导入对番茄乙烯生成的抑制作用，华中农业大学学报，1996，15(4)：305～309
[106] 王春霞，简志英，ACC合成酶基因及其反义基因对西瓜的遗传转化，Acta Botanica Siniea(植物学报：英文版)，1997，39(5)：445～450
[107] 陈锦清，郎春秀，反义PEP基因调控油菜籽粒蛋白质／油脂含量比率的研究，农业生物技术学报，1999，7(4)：316～320
[108] 郊娥香，杨志行等，水稻bicoid反义基因转基因植株胚胎发育的形态变化，植物学通报，2001，18(6)：707～713
[109] Fire A, Xu S, Montgomery MK, Kostas SA, et al, Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans., Nature, 1998, 391 (6669): 806～811
[110] Guo S, Kemphues KJ., par-1, a gene required for establishing polarity in C. elegans embryos, encodes a putative Ser/Thr kinase that is asymmetrically distributed, Cell. 1995 May 19; 81(4): 611-20. Cell. 1995 May 19; 81(4): 611～20
[111] 马鹏鹏，薛社普，韩代书，RNA干扰技术的原理与应用，中国组织化学与细胞化学杂志，2003，12(2)：201～207
[112] Brantl S., Antisense-RNA regulation and RNA interference, Biochim Biophys Acta. 2002 May3; 1575(1-3): 15～25
[113] Colaiacovo MP, Stanfield GM, Reddy KC, Reinke V, Kim SK, Villeneuve AM., A targeted RNAi screen for genes involved in chromosome morphogenesis and nuclear organization in the Caenorhabditis elegans germline, Genetics. 2002 Sep; 162(1): 113～28
[114] Yu JY, DeRuiter SL, Turner DL., RNA interference by expression of short-interfering RNAs and hairpin RNAs in mammalian cells, Proc Natl Acad Sci U S A. 2002 Apr 30; 99(9): 6047～6052.
[115] Czaudema F, Fechtner M, Aygun H, Arnold W, Klippel A, Giese K, Kaufmann J., Functional studies of the PI(3)-kinase signalling pathway employing synthetic and expressed siRNA.Nucleic Acids Res. 2003 Jan 15; 31(2): 670～682.
[116] 朱晓燕，刘宇健等，利用载体介导的RNA干扰技术抑制小鼠腹腔巨噬细胞RAW264．7内糖皮质激素受体表达及功能的研究，中国病理生理杂志，2003，19(11)：1541～1542
[117] Yu Shu-wen, Tang Zhang-cheng: Plant Physiology and Molecular Biology (2ed). Science Press. 1998.
[118] Forde, J., Malpica, J.M., The nucleotide sequence of a HMW glutenin subunit gene located on chromosome 1A of wheat (Triticum aestivum L.), Nucleic Acids Res. 13 (19), 6817-6832 (1985)
[119] Gu, Y.Q., Coleman-Derr, D.A., Rapid Genome Evolution Revealed By Comparative Analysis Of Orthologous Regions From Four Triticeae Genomes
[120] Chapman, J., Direct Submission(GeneBank), Apr 11, 2003
[121] Bhairi S M, Staples R C. 1992. Transient expression of the β-Glucuronidase Gene introduced into Uromyces appendicμlatus uredospores by particle bombardment. Phytopathology. 82(9):986～989
[122] Chen X, Stone M, Schlagnhaufer C, Romaine C P. 2000. A fruiting body tissue method for efficient Agrobacterium mediated transformation of Agaricus bisporus. Applied and Environmental Microbiology. 66(10):4510～4513
[123] Smith CJ, Watson CF, Morris PC, et, al, Inheritance and effect on ripening of antisense polygalacturonase genes in transgenic tomatoes, Plant Mol Biol. 1990 Mar: 14(3) :369-79
[124] Tentler S, Palas J, Enderlin C, et al. 1997. Inhibition of Neurospora crassa growth by a glucan synthaser-1 antisense construct. Current Microbiology. 34:303～308
[125] (美)J．萨姆布鲁克等，《分子克隆》第三版，2002：96～99，103～105，26～35