| 苏云金芽孢杆菌S1478—1分离株的特性鉴定及cry1Ac同源基因克隆 |
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| 引用本文： | 张文飞,姚淑霞,谢柳,杨明坤,王锐萍,叶大维,方宣钧.苏云金芽孢杆菌S1478—1分离株的特性鉴定及cry1Ac同源基因克隆[J].广西农业生物科学,2009(3):471-476. |
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| 作者姓名： | 张文飞 姚淑霞 谢柳 杨明坤 王锐萍 叶大维 方宣钧 |
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| 作者单位： | 广西大学生命科学与技术学院;海南海德热带农业资源研究所;海南师范大学生命科学院;南开大学生命科学院; |
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| 基金项目： | 中国Bt资源收鉴与利用项目;;国家863项目(2006AA022189)共同资助 |
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| 摘 要： | 本研究对从海南岛尖峰岭热带雨林自然保护区的土壤样品中分离出的＆菌株S1478～1进行了特性鉴定，研究表明S1478—1分离株菌落形态和生长特征和励参照菌株HD73极其相似。16SrDNA序列分析表明，S1478—1分离株与其它＆thuringiensis、B．cereus和B.anthracis的16S rDNA序列相似性达到99％。分离株能产菱形伴胞晶体，SDS-PAGE蛋白电泳分析表明，菌株在生长后期，形成芽孢同时分泌130kD大小的晶体蛋白。生物测定表明S1478—1分离株对小菜蛾具有很高的毒杀活性，LC50值高达5．159×10^8cfu／mL。初步显示S1478—1分离株可作为防治鳞翅目害虫的生物农药菌株。利用PCR—RFLP方法鉴定S1478—1分离株含有cry1Ac同源基因，以PCR粘性端克隆方法扩增全长基因，序列测定表明该基因ORF为3537bp，编码1178个氨基酸，推定的编码蛋白分子量为133．3kD，与其它cry1Ac基因序列最高达到99％同源，因此，该基因可作为杀虫工程菌及培育转基因抗虫作物的候选基因。
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| 关 键 词： | 苏云金芽孢杆菌 S1478—1分离株 cry1Ac同源基因 生物测定 基因登录号GQ227507 |
Identification of Bacillus thuringiensis Isolate S1478-1 and Cloning of cry1Ac Homolog |
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| Zhang Wenfei , Yao Shuxia Xie Liu , Yang Mingkun Wang Ruiping Ye Dawei Fang Xuanjun College of Life Science , Technology,Guangxi University,Nanning, College of Life Sciences,Hainan Normal University,Haikou, College of Life Sciences,Nankai University,Tianjin,Haide Institute of Tropical Agricultural Resources,Sanya.Identification of Bacillus thuringiensis Isolate S1478-1 and Cloning of cry1Ac Homolog[J].Journal of Guangxi Agricultural and Biological Science,2009(3):471-476. |
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| Authors: | Zhang Wenfei Yao Shuxia Xie Liu Yang Mingkun Wang Ruiping Ye Dawei Fang Xuanjun College of Life Science Technology Guangxi University Nanning College of Life Sciences Hainan Normal University Haikou College of Life Sciences Nankai University Tianjin Haide Institute of Tropical Agricultural Resources Sanya |
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| Institution: | Zhang Wenfei 1,4 Yao Shuxia 1 Xie Liu 1,4 Yang Mingkun 2 Wang Ruiping 2 Ye Dawei 3 Fang Xuanjun 4 1 College of Life Science , Technology,Guangxi University,Nanning,530005,2 College of Life Sciences,Hainan Normal University,Haikou,570200,3 College of Life Sciences,Nankai University,Tianjin,300071,4Haide Institute of Tropical Agricultural Resources,Sanya,572025 |
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| Abstract: | The identifications of a Bacillus thuringiensis isolate S1478-1, collected from the Jianfengling tropical rain forest reserves in Hainan Island, were carried out in the research.The results showed that the colonial mor-phology and growth curve of Bt isolate S1478-1 have quite similar with that of the standard strain kurstaki HD73.16S rDNA conserved sequence of Bt isolate S1478-1 was sequenced and its blast analysis of the sequence revealed that the Bt isolate S1478-1 should have high similarity compared wit... |
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| Keywords: | Bacillus thuringiensis S1478-1 isolate cry1Ac homolog Bioassay |
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