| Bt4.0718 cry1Ac基因的克隆与表达分析 |
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| 引用本文： | 丁学知,张何,孙运军,黄潢,张春艳,夏立秋.Bt4.0718 cry1Ac基因的克隆与表达分析[J].激光生物学报,2006,15(6):563-570. |
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| 作者姓名： | 丁学知 张何 孙运军 黄潢 张春艳 夏立秋 |
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| 作者单位： | 湖南师范大学生命科学学院,湖南,长沙,410081 |
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| 基金项目： | 国家自然科学基金;湖南省自然科学基金 |
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| 摘 要： | 在基因库中比对14种cry1Ac基因序列,发现了同源性很高的上游启动子区域和下游终止子区域。根据这一同源序列设计引物,从B t4.0718中扩增出包含双启动子和终止子的4.2 kb片段,用PCR-RFLP检测确定其中含有cry1Ac基因。然后将此片段克隆到穿梭载体pHT304中,转化大肠杆菌DH5α和B t无晶体突变株XZM-101。同时,利用原子力显微镜观察发现重组菌株BXZM34能够产生菱形晶体。
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| 关 键 词： | 苏云金芽孢杆菌 cry1Ac基因 穿梭载体 重组菌株 |
| 文章编号： | 1007-7146(2006)06-0563-08 |
| 收稿时间： | 2006/5/20 |
| 修稿时间： | 2006年5月20日 |
Cloning and Expression Analysis of the cry1Ac Gene from Bacillus thuringiensis Strain 4.0718 |
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| DING Xue-zhi,ZHANG He,SUN Yun-jun,HUANG Huang,ZHANG Chun-yan,XIA Li-qiu.Cloning and Expression Analysis of the cry1Ac Gene from Bacillus thuringiensis Strain 4.0718[J].ACTA Laser Biology Sinica,2006,15(6):563-570. |
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| Authors: | DING Xue-zhi ZHANG He SUN Yun-jun HUANG Huang ZHANG Chun-yan XIA Li-qiu |
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| Abstract: | 14 cry1Ac genes from GenBank were aligned and the consensus regions in the upstream of promoter and in the downstream of terminator were found. Based on the consensus sequences, a pair of primers was designed and a 4.2 kb element was amplified that includes the dual overlapping promoter and the whole termination-associated sequence from Bacillus thuringiensis strain 4. 0718, and the amplified 4.2 kb element was confirmed to contain the crylAc gene by cryI sub-genetype PCR-RFLP cry gene typing system. The 4.2 kb element was cloned into Bt-E. coli shuttle vector pHT30.4 and the cry1Ac gene was also expressed in E. coli DH5α and acrystalliferous mutant XZM-101. Meanwhile,rhombic crystal was observed from recombinant strain BXZM34 by atomic force microscope. |
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| Keywords: | Bacillus thuringiensis cry1Ac gene shuttle vector recombinant strain |
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