| B.t.c.cry 1C全长基因的克隆及其在增产菌中的表达 |
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| 引用本文： | 陈月华,李红秀,王津红,蔡峻,任改新. B.t.c.cry 1C全长基因的克隆及其在增产菌中的表达[J]. 微生物学通报, 2004, 31(1): 65-68 |
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| 作者姓名： | 陈月华 李红秀 王津红 蔡峻 任改新 |
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| 作者单位： | 1. 南开大学生命科学学院,天津,300071
2. 南开大学生命科学学院,天津,300071;天津农业生物技术研究中心,天津,300192 |
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| 基金项目： | 天津市科委,天津市农业生物技术研究中心科技(攻关)资助项目(No.033122111-1)天津市自然科学基金项目资助(No.023608811) |
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| 摘 要： | 设计的一对引物，对苏云金芽孢杆菌科默尔亚种(Bacillus thuringiensis subsp．colmeri)15A3菌株中的cry1C基因进行PER扩增，得到包括结构基因，调节基因在内的全长为4．0kb的PCR产物。经两步克隆，将此基因连接至穿梭表达载体pHT315上，得到重组质粒pHT-1C。通过电转化将其导入一株增产菌蜡状芽孢杆菌(Bacillus cereus)9509菌株，SDS-PAGE检测到1条60kD左右的蛋白带，镜检观察到菱形晶体，生测结果表明，cry1C基因的导人使Bc9509菌株获得了对甜菜夜蛾的杀虫活性。
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| 关 键 词： | 苏云金芽孢杆菌默尔亚种 cry1C 增产菌 杀虫活性 |
| 文章编号： | 0253-2654(2003)01-0065-04 |
| 修稿时间： | 2003-01-24 |
CLONING OF FULL CRY1 C GENE FROM B.T.C.AND EXPRESSION IN CROPS BENEFICIAL BACTERIA BACILLUS CEREUS |
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| CHEN Yue-Hua LI Hong-Xiu WANG Jin-Hong CAI Jun REN Gai-Xin. CLONING OF FULL CRY1 C GENE FROM B.T.C.AND EXPRESSION IN CROPS BENEFICIAL BACTERIA BACILLUS CEREUS[J]. Microbiology China, 2004, 31(1): 65-68 |
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| Authors: | CHEN Yue-Hua LI Hong-Xiu WANG Jin-Hong CAI Jun REN Gai-Xin |
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| Abstract: | Specific primers 1CaA/1CaB for full cry1C gene in Bacillus thuringiensis subsp.colmeri strain 15A3 were designed. The 4.0kb PCR product included the whole ORF and regulation region of cry1C gene. This PCR product was linked with shuttle vector pHT315 by two cloning steps. The recombined plasmid pHT-1C was electroporated into Bacillus cereus 9509, a kind of bacteria that beneficial to crops. The transformant could produce bipyramidal-shaped parasporal inclusions. The 60kD protein band was detected by SDS-PAGE. The bioassay result showed that the cry1C gene transformant of Bc 9509 had insecticidal activity to Spodoptera exigua. |
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| Keywords: | Bacillus thuringiensis subsp.colmeri cry1C Bacillus cereus Spodoptera exigua |
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