| B.thuringiensis穿梭载体的构建及cry1C基因的表达 |
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| 引用本文： | 何翔,刘坤,李冬颖,马明,耿运琪,陈启民.B.thuringiensis穿梭载体的构建及cry1C基因的表达[J].遗传学报,2000,27(7):647-653. |
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| 作者姓名： | 何翔 刘坤 李冬颖 马明 耿运琪 陈启民 |
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| 作者单位： | 南开大学生命科学学院，天津 300071 |
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| 基金项目： | 天津自然科学基金（973607511）；国家“九五”攻关项目（96-C01-0201）资助 |
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| 摘 要： | 以ＵＣ１９为母体，克隆了Ｂｔ ｋｅｎ－Ａｇ（Ｂ。．ｔｈｒｕｉｎｇｉｅｎｓｉｓ ｓｕｂｓｐ．ｋｅｎｙａｅ Ａｇ）的复制起始区（～１．６ｋｂ）、ｐＵＣ４Ｋ的ａｐｈ１基因，构建成穿梭载体ｐＨＶ－１，ｐＨＶ－１在Ｅ．ｃｏｌｉ中经１００个世代，质粒保持率在８０％以Ｂｔｉ ４Ｑ８（Ｂ．ｔｈｕｒｉｎｇｉｅｎｓｉｓ ｓｕｂｓｐ．ｉｓｒａｅｌｅｎｓｉｓ ４Ｑ８）中经４０个世代，质粒保持率在８０％以上，将Ｂ．ｌｉｃｈ
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| 关 键 词： | 穿梭载体 Bt cry1C基因 生物杀虫剂 遗传改良 |
| 文章编号： | 0379-172（2000）07-647-7 |
| 修稿时间： | 1999年9月10日 |
Construction of B. thuringiensis Shuttle Vector and Expression of the cry1C Gene |
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| HE Xiang,LIU Kun,LI Dong-Ying,MA Ming,GENG Yun-Qi,CHEN Qi-min.Construction of B. thuringiensis Shuttle Vector and Expression of the cry1C Gene[J].Journal of Genetics and Genomics,2000,27(7):647-653. |
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| Authors: | HE Xiang LIU Kun LI Dong-Ying MA Ming GENG Yun-Qi CHEN Qi-min |
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| Abstract: | We have constructed the E. coli-Bt shuttle vector pHV-1 by cloning the replicon (～1.6kb) of Bt ken-Ag and the aphI gene of pUC4K into pUC19. The rate of plasmid maintenance is more than 80% after 100 generations in E. coli, whereas 80% after 40 generations in Bti 4Q8. We have also constructed pHV-cry1C through cloning the α-mylase promoter from B. licheniformis and the cry1C gene from Bt 9510 into pHV-1 and introduced it into Bti 4Q8 by means of electroporation. Under the microscope, we can see that there is no crystal in Bti 4Q8, however, there are many rhomboid crystals in Bti 4Q8 (pHV-cry1C), which are smaller than those of Bt 9510. The bioassay result of Bti 4Q8 (pHV-cry1C) demonstrates that the expressed crystal protein is insecticidally active against Spodoptera exigue. |
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| Keywords: | shuttle vector Bacillus thuringiensis cry1C gene expression |
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