抗氧化酶类和耐辐射奇球菌抗紫外辐射的研究
|
摘要
超氧化物歧化酶(SOD)是一类有效的自由基清除剂。本研究通过检测SOD对溶原菌λ噬菌体紫外诱导的影响，探讨SOD对细菌紫外辐射的保护作用，确定λ原噬菌体紫外诱导过程中产生自由基的主要类型。结果表明加入SOD能明显地抑制λ噬菌体紫外诱导，其诱导效率与SOD浓度相关。当细菌悬浮液中SOD浓度为50u／mL时，产生的抑制效果最高(约为50)，发现氧化氢酶CAT抑制紫外诱导能力很弱，仅为10.7％左右，如果同时加入SOD和CAT，则能大大地提高溶原细胞紫外辐射的抑制率(72％)。综合本实验室以前的研究结果，表明λ溶原菌在紫外诱导过程中产生多种类型的自由基，其中以O_2~(．-)和·OH为主，并伴随有H_2O_2的形成。
通过PCR扩增和DNA重组技术，构建含有来自Bacillus stubtitls SOD基因的重组质粒pUC-SOD3，导入λ溶原菌后提取转化子质粒DNA，通过酶切分析、Southern杂交等方法证明SOD基因整合在pUC18上，并在溶原菌E.coli(λ)有效表达，表现出与外加SOD最佳效果的抗辐射能力。
应用电子自旋共振法及其自旋捕捉技术，对λ原噬菌体紫外诱导过程中的自由基进行直接检测，在电子顺磁共振仪显示屏上出现很强的自由基信号。加入SOD后，自由基的信号明显减弱。说明在λ原噬菌体紫外诱导过程中产生了自由基，而且SOD能有效的消除这些自由基。
此外，对耐辐射奇球菌(Deinococcus radiodurans)的抗紫外辐射特性进行检测。通过不同的方法从这种细菌细胞中提取抗辐射物质，应用HPLC、紫外扫描和显色反应初步认为这种抗辐射物质为类胡萝卜素。
通过分水岭算法能有效地将连在一起的菌落从电子图像中分割开，使平板菌落自动计数获得应用。该方法快速简便、准确、可靠，也适合于平板噬菌斑自动计数。
Superoxide dismutase (SOD) is an effective scavenger of free radicals. In order to determinate the protection of SOD, and the major species of free radicals, the effects of SOD were tested in the suspension of lysogenic bacteria by ultraviolet irradiation. The results indicated that SOD could significantly inhibit UV-induced of λ. prophage with a maximal inhibitory rate of 50%, and that the longer the UV-irradiated, the lower the inhibitory rate was. Meanwhile, it was found that CAT could inhibit UV-induced of A prophage with a maximal inhibitory rate of 10.7%. However, the mixture of CAT and SOD could greatly increase their inhibitory effect with a maximal inhibitory rate of 72%. In the present paper, free radicals species were analysed. It suggests that the complex free radicals were generated in the UV-induced of λ. prophage. However, 02 , ?OH and H2O2 played a dominant role in the induction.
Using the techniques of PCR and DNA recombination, a recombinant plasmid carrying SOD gene from Bacillus subtilis was constructed. The plasmid was transformed into the lysogenic bacteria E. coli ( A ). With analysis of DNA fragment of recombination plasmid, Southern bloting, the results indicated that SOD gene was ligated to the plasmid pUC18. The gene could effectively expressed in lysogenic bacteria. The recombinant strain could protect itself from UV-irradiated as SOD added in the suspension of lysogenic bacteria.
By electron spin resonance (ESR) and spin-trapping technique, the free radicals were detected in the suspension of the λ. lysogenic bacteria. The signals of free radical were observed when the suspension was irradiated by ultraviolet light. However, the main signals were disappeared in the suspension in the presence of SOD. It indicted that free radicals were generated in the process of UV-induced of λ. lysogenic bacteria, and that SOD could efficiently quench the free radicals.
Deinococcus radiodurans was also investigated in present study. It was found that a substance extracted from Deinococcus radiodurans by different methods, could protect cells from UV-irradiated. With analysis of HPLC, ultraviolet scanning and colorization, the radiation-resistant substance was identified as carotenoid.
With watershed algorithm, the linked colonies in the electrical images, which were prepared by scanning, were separated each other. Therefore, The colonies counting become an automatic method by computer. The method is a rapid, accurate, and is also suit to plaques counting on the plate.
通过PCR扩增和DNA重组技术，构建含有来自Bacillus stubtitls SOD基因的重组质粒pUC-SOD3，导入λ溶原菌后提取转化子质粒DNA，通过酶切分析、Southern杂交等方法证明SOD基因整合在pUC18上，并在溶原菌E.coli(λ)有效表达，表现出与外加SOD最佳效果的抗辐射能力。
应用电子自旋共振法及其自旋捕捉技术，对λ原噬菌体紫外诱导过程中的自由基进行直接检测，在电子顺磁共振仪显示屏上出现很强的自由基信号。加入SOD后，自由基的信号明显减弱。说明在λ原噬菌体紫外诱导过程中产生了自由基，而且SOD能有效的消除这些自由基。
此外，对耐辐射奇球菌(Deinococcus radiodurans)的抗紫外辐射特性进行检测。通过不同的方法从这种细菌细胞中提取抗辐射物质，应用HPLC、紫外扫描和显色反应初步认为这种抗辐射物质为类胡萝卜素。
通过分水岭算法能有效地将连在一起的菌落从电子图像中分割开，使平板菌落自动计数获得应用。该方法快速简便、准确、可靠，也适合于平板噬菌斑自动计数。
Superoxide dismutase (SOD) is an effective scavenger of free radicals. In order to determinate the protection of SOD, and the major species of free radicals, the effects of SOD were tested in the suspension of lysogenic bacteria by ultraviolet irradiation. The results indicated that SOD could significantly inhibit UV-induced of λ. prophage with a maximal inhibitory rate of 50%, and that the longer the UV-irradiated, the lower the inhibitory rate was. Meanwhile, it was found that CAT could inhibit UV-induced of A prophage with a maximal inhibitory rate of 10.7%. However, the mixture of CAT and SOD could greatly increase their inhibitory effect with a maximal inhibitory rate of 72%. In the present paper, free radicals species were analysed. It suggests that the complex free radicals were generated in the UV-induced of λ. prophage. However, 02 , ?OH and H2O2 played a dominant role in the induction.
Using the techniques of PCR and DNA recombination, a recombinant plasmid carrying SOD gene from Bacillus subtilis was constructed. The plasmid was transformed into the lysogenic bacteria E. coli ( A ). With analysis of DNA fragment of recombination plasmid, Southern bloting, the results indicated that SOD gene was ligated to the plasmid pUC18. The gene could effectively expressed in lysogenic bacteria. The recombinant strain could protect itself from UV-irradiated as SOD added in the suspension of lysogenic bacteria.
By electron spin resonance (ESR) and spin-trapping technique, the free radicals were detected in the suspension of the λ. lysogenic bacteria. The signals of free radical were observed when the suspension was irradiated by ultraviolet light. However, the main signals were disappeared in the suspension in the presence of SOD. It indicted that free radicals were generated in the process of UV-induced of λ. lysogenic bacteria, and that SOD could efficiently quench the free radicals.
Deinococcus radiodurans was also investigated in present study. It was found that a substance extracted from Deinococcus radiodurans by different methods, could protect cells from UV-irradiated. With analysis of HPLC, ultraviolet scanning and colorization, the radiation-resistant substance was identified as carotenoid.
With watershed algorithm, the linked colonies in the electrical images, which were prepared by scanning, were separated each other. Therefore, The colonies counting become an automatic method by computer. The method is a rapid, accurate, and is also suit to plaques counting on the plate.
引文
马兆勉、 陶纯堪。 自然背景下人工目标的检测与分割。 中国激光，2000，27(3) ：237-242
王惠媛。国际自由基生物学研讨会情况介绍。生命的化学，1988，8(6) ：37-45
王积分、赵增华、舒炎泰。小波分析用于菌落图像的纹理分割。计算机研究和发展，1999，36(12) ：237-242
孙存普、张建中、段介瑾。 自由基生物学导论。合肥： 中国科学技术大学出版社，1999，p66-81
朱文杰、洪燕萍、黄宁、蔡在龙。细菌SOD对微生物紫外光辐射损伤的恢复作用。应用与环境微生物学报，1996，2(1) ：79-83
华跃进、高冠军。耐辐射异常球菌DNA的损伤与修复相关基因的比较基因组研究。微生物学报，2003，43(1) ：120-127
张建中。 自旋标记ESR波谱的基本理论和应用。北京：科学出版社，1987，p353-359
张泽龙、马书林。SOD模拟物的研究进展。药学学报，2003，37(3) ：235—240
张超、刘军。SPC生产菌海洋红酵母摇瓶培养的研究。四川轻工学院学报，2000，13(3) ：71-75
杜泽吉、高桥佳子、鸣海一城。抗辐射Deinococcus radiodurans的生长特征及形态特征。苏州医学院院报，1998，18(11) ：1131-1132
宋道军、张立宏、余增亮。N~+离子注入对不同辐射敏感性微生物SOD，CAT和POD活性的影响。生物物理学报，1998，14(2) ：325-329，
宋道军、余增亮。三种辐射源对耐辐射微球菌作用机理的比较研究。生物物理学报，1998，14(1) ：185-188
宋道军、高冠军。耐辐射异常球菌抗辐射机理的研究进展。生命科学，1999，11(5) ：222—224
周士新。超氧化物歧化酶及其金属辅基与恶性肿瘤的关系。国外医学卫生学分册，1995，22(4) ：228—233
林克椿、吴本阶。医学生物物理。北京：北京医科大学，中国协和医科大学联合出版社，1996，p446—448
杨婉身、苟琳、张启军、于迟力。铜锌离子对超氧化物歧化酶构象及催化活性的影响。四川农业大学学报，1997，15(1) ： 138—140
杨雄。超氧化物歧化酶的研究现状。武汉教育学院学报，1999，18(3) ：49—54
赵保路。氧自由基和天然抗氧化剂。科学出版社，1999，p54—55
陆建峰、杨静宇、唐振民。重叠细胞图像分离算法的设计。计算机研究和发展，2000，37(2) ：112-117
陈小梅、倪国强、刘明奇。基于分水岭算法的红外图像分割方法。光电子-激光，2001，10(12) ：1069—1075．
范成鹏、方呈样、张骆珍、戴淑香。测定λ原噬菌体诱导频率的新方法。中国病毒学，2002，17(4) ：367—370
袁勤生。超氧化物歧化酶活性中心铜离子的还原氧化研究。生物化学杂志，1991，7(2) ：27—35
郝常明、王德培、丁友昉。光合细菌中类胡萝卜素德提取及性质德研究。天津轻工业学院院报，1999，3：5—11
黄建、 刘建国。 类胡萝卜素的分析测定。 青岛化工学院院报，2001，22(3) ：25—32
魏俊发、俞贤达、金道森。锰超氧化物歧化酶及其化学模拟的研究。化学进展，1997，9(1) ：14—21
Andley U P, Clark B A. The effects of near UV-radiation on human lens beta-crytallins: protein structural changes and the production of O_2 and H_2O_2. Photochem Photobiol., 1989, 50: 97
Anderson A W, Nordan H C, Cain R F. Studies on a radio resistant micrococcus RI, isolation, morphology, cultural characteristics and resistance to gamma radiation. Food Technol., 1956, 10:575-578
Bernard B K, McCord J M, Fridovich I. Superoxide dismutase from Esherichia coli B. J Boil Chem., 1970, 245 (22) : 6176-6181
Bieniek A, Moga A. An efficient watershed algorithm based on connects components. Pattern Recognition, 2000, 3: 907-916
Cohen S N, Chang A C Y, Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sic, 1972, 69:2110-2114
Chang L, Slot W, Geuze H J. Molecular immunocyto chemistry of the CuZn-superoxide dismutase in rat hepatocytes. J Cell Biol., 1988, 107 (6) : 2169-2175
Chou F I, Tans T. Mn~(2+) induction cell division and increasing in SOD and CAT activities in an aging Deinococcus radiodurans Cuture. J Bacteriol., 1990, 4:2029-2035
Daly M J, Long OY, Kenneth W M. Interplasmidic Recombination following irradiation of the radioresistant bacterium Deinococcus radiodurans. J Bacteriol., 1994, 7506-7551
Daly M J, Kenneth W M. An alternative pathway of recombination of chromosomal fragment precedes recA-dependent recombination in the radioresistant bacterium Deinococcus radiodurans. J Bacteriol., 1996, 4461-4471.
Daly M J, Kenneth W M. Recombination between a resident plamid and in the chromosome following irradiation of the radioresistant bacterium Deinococcus radiodurans. Gene, 1997, 187:225-229
Guyton K, Kensler T W. Oxidative mechanisms in carcinogenesis. Brit. Med. bull., 1993, 49:523-544
Hass H M, Massaro D. Differences in Cu, Zn-SOD induction in lungs of neonatal and adult rats. Am J Physiol., 1987, 235:66-72
Halliwell B. Free radicals and antioxidants: a personal view. Nutr. Rev., 1994, 52:253-265
Hansen W M, Williams E H. Watershed-based maximum-homogeneity filtering. IEEE Trans, 1999, 8(7) : 982-988
Igor N Z, Thomas J M. Superoxide dismutase multigene family: a comparison of the Cu, Zn-sod (sod1) , Mn-sod (sod2) , and Fe-sod (sod3) gene structures, evolution and expression. Free radical biology & medicine, 2002, 33(3) : 337-349
Jachson S M. An analysis of structural similarity in the iron and manganese superoxide dismutase based on known structures and sequences. Biometals, 1998, 11:159—173
John R B, Ashlee M E, Park M J. Why is Deinococcus radiodurans so resistant to ionizing radiation. Trends in Microbiology, 1999, 7(9) : 362-364
Jeffrey K, Reddy R, Luke G. Effects of haloperidol on antioxidant defense system enzymes in schizophrenia. J Psychiatric Res., 1998, 32:385-394
John R B. Radiation rasistance: the fragments that remain. Current biology. 2000, 10: 204-205,
Kenneth W M. Repair of ionizing-radiation damage in the radiation resistant bacterium Deinococcus radiodurans. Mutation Research, 1996, 36(3) : 1-7
Kira S M, Aravind L Y. Genome of the extremely radiation-resistant Bacterium Deinococcus radiodurans viewed from the perspective of comparative genomic. Microbiology and Molecular Biology Reviews, 2001, 65(1) : 44-79
Kira S M, Aravind L Y. Short repeats and IS elements in the extremely radiation-resistant bacterium Deinococcus radiodurans. Microbiol Res., 1999, 150:711-724
Keller G A, Warnes T G, Steiner K S. Cu, Zn-superoxide dismutase is a peroxisomal enzyme in human fibrobcasts and hepatoma cell. Proc Natl Acad Sci., 1991, 88(16) :7381-7390
Lemee L, Peuchant E, Clerc M. Deinoxanthin: a new carotenoid Isolate from Deinococcus radiodurans. Tetrahedron, 1998, 53(33) : 919-926
Lohr J B, Kuczenski R, Bracha H S. Increasing indices of free
radical activity in the cerebrospinal of patients with tardive dyskinesia. Biology Psychiatry, 1990, 28:535-539
Lohr J B. Oxygen radicals and neuropsychiatric illness:some speculation. Arch Gen Psychiatry, 1991, 48:1097-1106
Lin LY, Rong Q, Aston C. Whole-genome shotgun optical mapping of Deinococcus radiodurans. Science, 1999, 285:1558-1562
Lee JH, Choi I Y, Kil I S. Protective role of superoxide dismutases against ionizing radiation in yeast. Biochimi Biophysica Acta, 2001, 15(26) : 191-198
McCord J M, Fridovich J. Superoxide dismutase: an enmazymic function for erythrocuprein (hemocuprein) . J biol Chem., 1969, 22(25) : 6040-6055
Marklund S. Spectrophotometric study of spontaneous disproportination of superoxide anion radical and sensitive directassay for superoxide dismutase. J Biol Chem., 1976, 251(23) : 7504-7507
Mayer F, Beucher S. Morphology segmentation. Visual and Image Representation, 1990, 1:21-46
Metin O, Mehmet M, Cagatay O. Increased oxidative stress and hypozincemia in male obesity. Pans, 2002, 35:627-631.
Michhael B D, Harrop H A, Held K D. Photoreactivation of Escherichia coli after exposure to ionizing radiation: the role of UV damage by concomitant cerenkov light. International Journal of Radiation Biology, 1981, 39 (5) : 577-582
Murray N E, Murry K. Manipulation of restriction targetsin phage
λ to form receptor chromosomes for DNA fragment. Nature, 1974, 251:53-61
Madsen H L, Nielsen B R, Bertelsen G, Skibsted L H. Screening of antioxidative activities of speices, a comparison between assays based on ESR spin trapping and elecfrochemical measurement of oxygen consumption. Food chemistry, 1996, 2:331-337
Peak M J, Peak J G. Hydroxyl radical quenshing agents protece against DNA breakage caused by both 365nm UV-A and by gamme radication. Photochem Photobiol., 1990, 51:649-657
Smadar L Z, Joseph E M. Ringlike structor of the Deiococcus radioradurans genone: a key to radioresistance. Science, 2003, 299:224-226
Stefano G, Fabiana F, Elida F. Determination of superoxide dismutase in erythrocytes by a chemiluminescent assary. Talanta, 2000, 51:685-692
Sambrook J, David W R. Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press. 2001
Thomas M, Cameron F R, Davis R W. Viable molecular hybrids of bacteriophage lambda and eukaryotic DNA. Proc Natl Acad Sic., 1974, 71:4579
Totter J R. Spontaneous cancer and its possible relationship to oxygen metabolism, Proc Natl Acad Sic., 1980, 77:1763
Tainer J A. Detemination and analyzis of the struction of copper-zinc superoxide dismutase. J Mol Biol, 1982, 160:181-190
Takashi I, Yoshinobu M, Tetsuaki T. Molecular cloning and
nucleotide sequence of the superoxide dismutase gene and characterization of its product from Bacillus subtilis. J Bacteriol., 1998:3697-3703
Vincent L, Soille P. Watersheds in digital space: an efficient algorithm based on immersion simulation. IEEE Trans., 1991, 18(12) : 1163-1173
Vuchetich P J. Naphthalene-induced oxidative stress in rats and the protective effects of vitamin E succinct. Free Radical Biol Med., 1996, 21 (5) :577
Wang P, Schellhor H E, Induction of resistance to hydrogen peroxide and radiation in Deinococcus radiodurans. Can J Mircobiol., 1995, 41:170-176
White O, Jonatha A, Eisen F. Genome sequence of the radioresistant bacterium Deinococcus radiodurans R1. Science, 1999, 286.
Liu YQ, Zhou JZ, Marina V O. Transcriptome dynamic of Deinococcus radiodurand recovering from ioning radiation. Pans, 2003, 100(7) : 4191-4196
Yost F, Fridovich I. An iron-containing superoxide dismutase from E. coli. J Biol Chem., 1973, 248:4905-4908
Ynch M L, Howard K T. Expression and role of superoxide dismutase in pathogenic bacteria. Microbes and infection, 2000, 2:1245-1255
Yoo S J, Young H K, Robert A. Copper, Zinc superoxide dismutase enhance DNA damage and mutagenicity induced by cysteine/iron. Mutation Research, 2000, 448:97-104
Zhu S H, Fang C X, Zhu S Q, Fan C P, Zhang L Z. Inhibitory effect
of gynostemma pentaphyllum on the UV induction of bacteraiophage λ in lysogenic Escherichia coli. Current Microbiology. 2001, 43: 299-301
王惠媛。国际自由基生物学研讨会情况介绍。生命的化学，1988，8(6) ：37-45
王积分、赵增华、舒炎泰。小波分析用于菌落图像的纹理分割。计算机研究和发展，1999，36(12) ：237-242
孙存普、张建中、段介瑾。 自由基生物学导论。合肥： 中国科学技术大学出版社，1999，p66-81
朱文杰、洪燕萍、黄宁、蔡在龙。细菌SOD对微生物紫外光辐射损伤的恢复作用。应用与环境微生物学报，1996，2(1) ：79-83
华跃进、高冠军。耐辐射异常球菌DNA的损伤与修复相关基因的比较基因组研究。微生物学报，2003，43(1) ：120-127
张建中。 自旋标记ESR波谱的基本理论和应用。北京：科学出版社，1987，p353-359
张泽龙、马书林。SOD模拟物的研究进展。药学学报，2003，37(3) ：235—240
张超、刘军。SPC生产菌海洋红酵母摇瓶培养的研究。四川轻工学院学报，2000，13(3) ：71-75
杜泽吉、高桥佳子、鸣海一城。抗辐射Deinococcus radiodurans的生长特征及形态特征。苏州医学院院报，1998，18(11) ：1131-1132
宋道军、张立宏、余增亮。N~+离子注入对不同辐射敏感性微生物SOD，CAT和POD活性的影响。生物物理学报，1998，14(2) ：325-329，
宋道军、余增亮。三种辐射源对耐辐射微球菌作用机理的比较研究。生物物理学报，1998，14(1) ：185-188
宋道军、高冠军。耐辐射异常球菌抗辐射机理的研究进展。生命科学，1999，11(5) ：222—224
周士新。超氧化物歧化酶及其金属辅基与恶性肿瘤的关系。国外医学卫生学分册，1995，22(4) ：228—233
林克椿、吴本阶。医学生物物理。北京：北京医科大学，中国协和医科大学联合出版社，1996，p446—448
杨婉身、苟琳、张启军、于迟力。铜锌离子对超氧化物歧化酶构象及催化活性的影响。四川农业大学学报，1997，15(1) ： 138—140
杨雄。超氧化物歧化酶的研究现状。武汉教育学院学报，1999，18(3) ：49—54
赵保路。氧自由基和天然抗氧化剂。科学出版社，1999，p54—55
陆建峰、杨静宇、唐振民。重叠细胞图像分离算法的设计。计算机研究和发展，2000，37(2) ：112-117
陈小梅、倪国强、刘明奇。基于分水岭算法的红外图像分割方法。光电子-激光，2001，10(12) ：1069—1075．
范成鹏、方呈样、张骆珍、戴淑香。测定λ原噬菌体诱导频率的新方法。中国病毒学，2002，17(4) ：367—370
袁勤生。超氧化物歧化酶活性中心铜离子的还原氧化研究。生物化学杂志，1991，7(2) ：27—35
郝常明、王德培、丁友昉。光合细菌中类胡萝卜素德提取及性质德研究。天津轻工业学院院报，1999，3：5—11
黄建、 刘建国。 类胡萝卜素的分析测定。 青岛化工学院院报，2001，22(3) ：25—32
魏俊发、俞贤达、金道森。锰超氧化物歧化酶及其化学模拟的研究。化学进展，1997，9(1) ：14—21
Andley U P, Clark B A. The effects of near UV-radiation on human lens beta-crytallins: protein structural changes and the production of O_2 and H_2O_2. Photochem Photobiol., 1989, 50: 97
Anderson A W, Nordan H C, Cain R F. Studies on a radio resistant micrococcus RI, isolation, morphology, cultural characteristics and resistance to gamma radiation. Food Technol., 1956, 10:575-578
Bernard B K, McCord J M, Fridovich I. Superoxide dismutase from Esherichia coli B. J Boil Chem., 1970, 245 (22) : 6176-6181
Bieniek A, Moga A. An efficient watershed algorithm based on connects components. Pattern Recognition, 2000, 3: 907-916
Cohen S N, Chang A C Y, Hsu L. Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sic, 1972, 69:2110-2114
Chang L, Slot W, Geuze H J. Molecular immunocyto chemistry of the CuZn-superoxide dismutase in rat hepatocytes. J Cell Biol., 1988, 107 (6) : 2169-2175
Chou F I, Tans T. Mn~(2+) induction cell division and increasing in SOD and CAT activities in an aging Deinococcus radiodurans Cuture. J Bacteriol., 1990, 4:2029-2035
Daly M J, Long OY, Kenneth W M. Interplasmidic Recombination following irradiation of the radioresistant bacterium Deinococcus radiodurans. J Bacteriol., 1994, 7506-7551
Daly M J, Kenneth W M. An alternative pathway of recombination of chromosomal fragment precedes recA-dependent recombination in the radioresistant bacterium Deinococcus radiodurans. J Bacteriol., 1996, 4461-4471.
Daly M J, Kenneth W M. Recombination between a resident plamid and in the chromosome following irradiation of the radioresistant bacterium Deinococcus radiodurans. Gene, 1997, 187:225-229
Guyton K, Kensler T W. Oxidative mechanisms in carcinogenesis. Brit. Med. bull., 1993, 49:523-544
Hass H M, Massaro D. Differences in Cu, Zn-SOD induction in lungs of neonatal and adult rats. Am J Physiol., 1987, 235:66-72
Halliwell B. Free radicals and antioxidants: a personal view. Nutr. Rev., 1994, 52:253-265
Hansen W M, Williams E H. Watershed-based maximum-homogeneity filtering. IEEE Trans, 1999, 8(7) : 982-988
Igor N Z, Thomas J M. Superoxide dismutase multigene family: a comparison of the Cu, Zn-sod (sod1) , Mn-sod (sod2) , and Fe-sod (sod3) gene structures, evolution and expression. Free radical biology & medicine, 2002, 33(3) : 337-349
Jachson S M. An analysis of structural similarity in the iron and manganese superoxide dismutase based on known structures and sequences. Biometals, 1998, 11:159—173
John R B, Ashlee M E, Park M J. Why is Deinococcus radiodurans so resistant to ionizing radiation. Trends in Microbiology, 1999, 7(9) : 362-364
Jeffrey K, Reddy R, Luke G. Effects of haloperidol on antioxidant defense system enzymes in schizophrenia. J Psychiatric Res., 1998, 32:385-394
John R B. Radiation rasistance: the fragments that remain. Current biology. 2000, 10: 204-205,
Kenneth W M. Repair of ionizing-radiation damage in the radiation resistant bacterium Deinococcus radiodurans. Mutation Research, 1996, 36(3) : 1-7
Kira S M, Aravind L Y. Genome of the extremely radiation-resistant Bacterium Deinococcus radiodurans viewed from the perspective of comparative genomic. Microbiology and Molecular Biology Reviews, 2001, 65(1) : 44-79
Kira S M, Aravind L Y. Short repeats and IS elements in the extremely radiation-resistant bacterium Deinococcus radiodurans. Microbiol Res., 1999, 150:711-724
Keller G A, Warnes T G, Steiner K S. Cu, Zn-superoxide dismutase is a peroxisomal enzyme in human fibrobcasts and hepatoma cell. Proc Natl Acad Sci., 1991, 88(16) :7381-7390
Lemee L, Peuchant E, Clerc M. Deinoxanthin: a new carotenoid Isolate from Deinococcus radiodurans. Tetrahedron, 1998, 53(33) : 919-926
Lohr J B, Kuczenski R, Bracha H S. Increasing indices of free
radical activity in the cerebrospinal of patients with tardive dyskinesia. Biology Psychiatry, 1990, 28:535-539
Lohr J B. Oxygen radicals and neuropsychiatric illness:some speculation. Arch Gen Psychiatry, 1991, 48:1097-1106
Lin LY, Rong Q, Aston C. Whole-genome shotgun optical mapping of Deinococcus radiodurans. Science, 1999, 285:1558-1562
Lee JH, Choi I Y, Kil I S. Protective role of superoxide dismutases against ionizing radiation in yeast. Biochimi Biophysica Acta, 2001, 15(26) : 191-198
McCord J M, Fridovich J. Superoxide dismutase: an enmazymic function for erythrocuprein (hemocuprein) . J biol Chem., 1969, 22(25) : 6040-6055
Marklund S. Spectrophotometric study of spontaneous disproportination of superoxide anion radical and sensitive directassay for superoxide dismutase. J Biol Chem., 1976, 251(23) : 7504-7507
Mayer F, Beucher S. Morphology segmentation. Visual and Image Representation, 1990, 1:21-46
Metin O, Mehmet M, Cagatay O. Increased oxidative stress and hypozincemia in male obesity. Pans, 2002, 35:627-631.
Michhael B D, Harrop H A, Held K D. Photoreactivation of Escherichia coli after exposure to ionizing radiation: the role of UV damage by concomitant cerenkov light. International Journal of Radiation Biology, 1981, 39 (5) : 577-582
Murray N E, Murry K. Manipulation of restriction targetsin phage
λ to form receptor chromosomes for DNA fragment. Nature, 1974, 251:53-61
Madsen H L, Nielsen B R, Bertelsen G, Skibsted L H. Screening of antioxidative activities of speices, a comparison between assays based on ESR spin trapping and elecfrochemical measurement of oxygen consumption. Food chemistry, 1996, 2:331-337
Peak M J, Peak J G. Hydroxyl radical quenshing agents protece against DNA breakage caused by both 365nm UV-A and by gamme radication. Photochem Photobiol., 1990, 51:649-657
Smadar L Z, Joseph E M. Ringlike structor of the Deiococcus radioradurans genone: a key to radioresistance. Science, 2003, 299:224-226
Stefano G, Fabiana F, Elida F. Determination of superoxide dismutase in erythrocytes by a chemiluminescent assary. Talanta, 2000, 51:685-692
Sambrook J, David W R. Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press. 2001
Thomas M, Cameron F R, Davis R W. Viable molecular hybrids of bacteriophage lambda and eukaryotic DNA. Proc Natl Acad Sic., 1974, 71:4579
Totter J R. Spontaneous cancer and its possible relationship to oxygen metabolism, Proc Natl Acad Sic., 1980, 77:1763
Tainer J A. Detemination and analyzis of the struction of copper-zinc superoxide dismutase. J Mol Biol, 1982, 160:181-190
Takashi I, Yoshinobu M, Tetsuaki T. Molecular cloning and
nucleotide sequence of the superoxide dismutase gene and characterization of its product from Bacillus subtilis. J Bacteriol., 1998:3697-3703
Vincent L, Soille P. Watersheds in digital space: an efficient algorithm based on immersion simulation. IEEE Trans., 1991, 18(12) : 1163-1173
Vuchetich P J. Naphthalene-induced oxidative stress in rats and the protective effects of vitamin E succinct. Free Radical Biol Med., 1996, 21 (5) :577
Wang P, Schellhor H E, Induction of resistance to hydrogen peroxide and radiation in Deinococcus radiodurans. Can J Mircobiol., 1995, 41:170-176
White O, Jonatha A, Eisen F. Genome sequence of the radioresistant bacterium Deinococcus radiodurans R1. Science, 1999, 286.
Liu YQ, Zhou JZ, Marina V O. Transcriptome dynamic of Deinococcus radiodurand recovering from ioning radiation. Pans, 2003, 100(7) : 4191-4196
Yost F, Fridovich I. An iron-containing superoxide dismutase from E. coli. J Biol Chem., 1973, 248:4905-4908
Ynch M L, Howard K T. Expression and role of superoxide dismutase in pathogenic bacteria. Microbes and infection, 2000, 2:1245-1255
Yoo S J, Young H K, Robert A. Copper, Zinc superoxide dismutase enhance DNA damage and mutagenicity induced by cysteine/iron. Mutation Research, 2000, 448:97-104
Zhu S H, Fang C X, Zhu S Q, Fan C P, Zhang L Z. Inhibitory effect
of gynostemma pentaphyllum on the UV induction of bacteraiophage λ in lysogenic Escherichia coli. Current Microbiology. 2001, 43: 299-301