| 大肠杆菌RNaseⅢ基因的克隆与表达 |
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| 引用本文： | 解岩,郭俊伟,赵兴卉,陈薇.大肠杆菌RNaseⅢ基因的克隆与表达[J].生物技术通讯,2008,19(4):532-534. |
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| 作者姓名： | 解岩 郭俊伟 赵兴卉 陈薇 |
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| 作者单位： | 北京微生物流行病研究所,病原微生物生物安全国家重点实验室(2004DAV00214) 北京,100071 |
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| 摘 要： | 目的：克隆并在原核表达系统中表达RNaseⅢ基因。方法：提取大肠杆菌JM109株的基因组DNA,以之为模板扩增得到RNaseⅢ基因全序列,将该编码序列克隆到原核表达载体pET-22b（＋）中,转化大肠杆菌BL21（DE3）,IPTG诱导重组RNaseⅢ表达。结果与结论：在大肠杆菌中克隆到了RNaseⅢ的全基因,经测序证明与数据库中报道的序列一致;表达的重组RNaseⅢ主要以包涵体形式存在。
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| 关 键 词： | RNaseⅢ 基因克隆 原核表达 |
Cloning and Expression of Escherichia coli RNaseⅢ |
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| XIE Yan,GUO Jun-Wei,ZHAO Xing-Hui,CHEN Wei.Cloning and Expression of Escherichia coli RNaseⅢ[J].Letters in Biotechnology,2008,19(4):532-534. |
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| Authors: | XIE Yan GUO Jun-Wei ZHAO Xing-Hui CHEN Wei |
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| Institution: | ( Beijing Institute of Microbiology and Epidemiology,State Key Laboratory of Pathgen and Biosecurity(2004DAV00214),Beijing 100071,China ) |
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| Abstract: | Objective：To clone and prokaryotic express the RNaseⅢ gene of Escherichia coli.Methods：The genomic DNA was extracted from E.coli strain JM109,and RNaseⅢ gene was amplified based on it,and was then inserted into a prokaryotic expression vector named pET-22b（＋）.The recombinant plasmids were transformed into E.coli BL21（DE3） and induced with IPTG.Results and Conclusion：The amplified RNaseⅢ gene was confirmed by DNA sequence analysis.The RNaseⅢ protein was successfully expressed in form of inclusion body. |
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| Keywords: | RNaseⅢ gene cloning prokaryotic expression |
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