DNA分子标记鉴定家蚕品种和纯度的研究与香樟核糖体失活蛋白的纯化
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摘要
依靠形态特征鉴定家蚕品种与纯度，易受环境影响，检验准确性差。本文应用DNA分子标记技术，利用RAPD特征和SCAR标记鉴定江苏省主要蚕品种及其纯度。结果找到了苏5、苏6，苏菊、明虎，苏镇、春光等品种的RAPD特征DNA片段，进一步将苏5、苏6，苏菊、明虎两组RAPD标记转换成特异性更强的SCAR标记，可以准确地用于苏5、苏6和苏菊、明虎品种及其后代的纯度鉴定。
核糖体失活蛋白(RIP)是生物体内存在的一类抑制真核生物核糖体蛋白质合成的蛋白质，可作为免疫毒素的毒弹头等，应用于治疗癌症和AIDS等多种疾病。本文对香樟Ⅱ型核糖体失活蛋白Cinnamomin进行了大规模的抽提和纯化。并将纯化的Cinnamomin蛋白与大鼠肝细胞80S核糖体作用，鉴定出其存在RNAN—糖苷酶的活性，确认其为核糖体失活蛋白。
The method of identifying silkworm variety and purity with morphological characters is easily affected by environment and the accuracy is poor.
In this paper, the DNA polymorphism bands have been amplified with RAPD among Jiangsu main silkworm races, Su5, Su6, Suju, Minghu, Suzhen, and Chunguang. Then respectively convert specific bands of Su5, Su6, Suju and Minghu into stable SCAR markers (Sequenced characterized amplified region).These SCAR markers can be used to determine variety of Su5, Su6, Suju and Minghu and the purity of their Fl hybrid.
Ribosome-inactivating proteins (RIPs) are a group of cytoxin which chemically act on eukaryotic ribosome rRNA and inhibit protein biosynthesis. Increasing interests concentrate in this field due to the potential use of RIPs for treatment of several important human diseases such as cancer and AIDS. Here, a new method for large-scale preparation of Cinnamomin, a RIP from Cinnamomum camphora was established. In this paper the activity of RNA N-glycosidas of the purified Cinnamomin was identified. Rat liver 80S ribosomes were incubated with Cinnamomin. The purified Cinnamomin is a kind of RIP.
核糖体失活蛋白(RIP)是生物体内存在的一类抑制真核生物核糖体蛋白质合成的蛋白质，可作为免疫毒素的毒弹头等，应用于治疗癌症和AIDS等多种疾病。本文对香樟Ⅱ型核糖体失活蛋白Cinnamomin进行了大规模的抽提和纯化。并将纯化的Cinnamomin蛋白与大鼠肝细胞80S核糖体作用，鉴定出其存在RNAN—糖苷酶的活性，确认其为核糖体失活蛋白。
The method of identifying silkworm variety and purity with morphological characters is easily affected by environment and the accuracy is poor.
In this paper, the DNA polymorphism bands have been amplified with RAPD among Jiangsu main silkworm races, Su5, Su6, Suju, Minghu, Suzhen, and Chunguang. Then respectively convert specific bands of Su5, Su6, Suju and Minghu into stable SCAR markers (Sequenced characterized amplified region).These SCAR markers can be used to determine variety of Su5, Su6, Suju and Minghu and the purity of their Fl hybrid.
Ribosome-inactivating proteins (RIPs) are a group of cytoxin which chemically act on eukaryotic ribosome rRNA and inhibit protein biosynthesis. Increasing interests concentrate in this field due to the potential use of RIPs for treatment of several important human diseases such as cancer and AIDS. Here, a new method for large-scale preparation of Cinnamomin, a RIP from Cinnamomum camphora was established. In this paper the activity of RNA N-glycosidas of the purified Cinnamomin was identified. Rat liver 80S ribosomes were incubated with Cinnamomin. The purified Cinnamomin is a kind of RIP.
引文
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2．周志康，李成仁，伍永瑜，夏庆友．家蚕的激光荧光特征与品种经济属性的饿相关性(J)．激光杂志，1991，12(6)：307～310
3. Avise J C. Molecular markers, natural history and evolution (M). New York: Chapman & Hall, Inc. 1994
4．陈景堂，池书敏，马占元，刘志增，魏俊杰等．玉米清蛋白聚丙烯酰胺电泳谱带编码与种子纯度检验(J)．华北农学报，20001S(增刊)：20～25
5．方玉春，张兴和，冯德举，刘俊平，庄炳昌，梁晓葵．应用盐蛋白电泳图谱鉴定玉米种子纯度(J)．吉林农业大学学报,2001，23(4)：6～10
6．潘良文．力口拿大谷物研究所品种鉴别研究简介(J)．现代商检科技，1998,8(6)：23～25
7．孙雁，冯锐，段利琴．杂交玉米及亲本种子碱溶蛋白电泳鉴定(J)．种子，2000，111(5)：17～18
8．石思信，张志娥，肖检平．利用蛋白质聚丙烯酰胺电泳法鉴定水稻种子纯度(J)．北京农业科学，1996，14(4)：8～10
9．余有海，张英，史桂琴，王弘．蛋白质凝胶电泳方法的几点改进．(J)．种子科技，2001，(3)：169
10．王晓峰，赵婷．超薄等电聚焦电泳技术及其在种子纯度鉴定中的运用(J)．种子，2000(6)：55～57
11．陈启林，王鸣，陈静．辣椒种子芽期同工酶动态变化与纯度鉴定关系探讨(J)．陕西农业科学，1996，(5)：11～13
12．郑晓鹰，黄为平．过氧化物酶及种子蛋白分析鉴定甜椒杂交种纯度(J)．华北农学报，1996，11(4)：119～122
13．陶芳，李旭，陈龙英，许国慰．酯酶同工酶谱分析鉴定杂交水稻种子纯度技术初探(J)．种子，1997，88(2)：55～57
14．王振英，陈宏，彭永康．用蛋白质和POD同工酶指纹图谱检测玉米自交系种子纯度．天津师大学报(自然科学版)，2000，20(4)：45～47
15．罗英，刘春，陈萍，代方银，鲁成．家蚕SOD酶品种间的差异研究(A)．中国蚕丝学会第三届青年学会材料，西南农业大学蚕桑丝绸学院(2001年10月，杭州)：129～136
16．霍绍棠，王炜，薛增召．三种赤眼蜂酯酶同工酶的比较研究(J)．西北农业学报，1994，3(1)：81～86
17．李成伟，何德银，李卓杰．用等电聚焦法对茄子种子几种同工酶的研究(J)．种子，1997，
92(6)：1～2
18．陈启林，王鸣，陈静．辣椒种子芽期同工酶动态变化与纯度鉴定关系探讨(J)．陕西农业科学，1996，(5)：11～13
19. Smith J V C, smith O .Restriction fragment ength polymorphisms can differentiate among US maize hybrids (J).Crop Science, 1991, 31:893～899
20．程道军，周泽扬，鲁成，向仲怀，曾华明，漆定梅，杨彪．RFLP技术构建家蚕现行DNA指纹图谱的研究(J)．西南农业大学学报，2000，22(6)：484～486
21. Amold M L, Bucker C M, Robinson J J. Pollen mediated introgression and hybrid speciation in Louisana irises (J).Pro Natl Acad Sci USA., 1991, 88:1398-1402
22. Liston A..Variation in the chroloplast genes rpo c1 and rpo c2 of thegenus Astragalus (Fabaceae) :Evidence from restriction site mapping of a PCR-amplified fragment (J). Amer J Bot., 1992,79:956-961
23. Rieseberg LH, Hanson M A and Philbrick CT.Andredioecy is derived from dioecy in Datiscaceae:Evidence from restriction site mapping of PCR-amplified chloroplast (J). Systmatic Botany., 1992,17 (2) :324-336
24. Mullis K B, Faloona F A, Schaof S et al. Specific amplification of DNA in vitro: the polymerase chain reaction (J). Cold Spring Harbor Smp Biol., 1986,51:263-273
25. Williams J G K, Kubelik A R, Livak K J. DNA polymorphism amplified by arbitrary primers are useful as gentic markers (J). Nucl Acids Res., 1990,18:6231-6235
26. Welsh J, McClelland M. Fingerprinting genomes using PCR with arbitrary primers (J). Nucl Acids Res., 1990,18:7213-7218
27. Caetano-Anolles G, Bassam B J, Gresshoff PM.DNA amLification fingerprinting using very short arbitrary oligonucleotide primers (J). Biotechnology, 9:553-557
28. Williams J G K, Hanafey M K, Rafalski J A et al. Genetic analysis using random amplified polymorthic DNA markers (J). Meth Enzymol., 1993, 218:704-740
29．赵久然，郭景伦，孔艳芳，池书敏，尉德铭，滕海涛．利用DNA指纹鉴定玉米杂交中纯度及其真伪技术的研究(J)．玉米科学，1999，7(1)：9-13
30．钟伯雄，丁农，张金卫，陶京亚，陆云等利用DNA指纹判断家蚕杂交纯度的方法(A)．中国蚕丝学会第三届青年学术讨论会．(2001年10月杭州)：83-85
31．翁宏飚，徐孟奎，张耀洲．家蚕的RAPD及其品种间的差异(J)．浙江农业大学学报，1996，22(2)：152-156
32. Novy R G, Vorsa N.Theor Appl Gene., 1996, 92:840
33．林万明等．PCR技术操作和应用指南(M)．人民军医出版社，1995
34．李培金．玉米RAPD影响因素及对生基因的分子标记研究：(硕士论文)．合肥：安徽农业大学．1999
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