| 肝素黄杆菌2-O-sulfatase基因的克隆与表达 |
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| 引用本文： | 余永莉,陈敬华,郑化.肝素黄杆菌2-O-sulfatase基因的克隆与表达[J].安徽农业科学,2011,39(12):7286-7288. |
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| 作者姓名： | 余永莉 陈敬华 郑化 |
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| 作者单位： | 余永莉,郑化(武汉理工大学化学工程学院,湖北武汉,430070);陈敬华(江南大学医药学院,江苏无锡,214122) |
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| 摘 要： | 目的]研究肝素黄杆菌2-O-sulfatase基因的克隆与表达。方法]以肝素黄肝菌为模板,通过PCR从肝素黄杆菌基因组DNA中扩增2-O-sulfatase的基因,测序正确后插入到表达质粒PET-28a（＋）上构建表达载体,重组质粒转化E.coil BL21（DE3）进行蛋白质表达。结果]成功地将PCR扩增得到的测序正确的2-O-sulfatase基因构建入PET-28a（＋）载体上,重组质粒转入E.coil BL21（DE3）后,表达产物经SDS-PAGE凝胶电泳鉴定得到目的蛋白。结论]克隆并成功表达了黄杆菌2-O-sulfatase基因,为分析肝素多糖分子及其降解产物结构奠定了基础。
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| 关 键 词： | 肝素黄杆菌 2-O-sulfatase 克隆 蛋白表达 纯化 |
Cloning and Expression of Flavobacterium heparinum 2-O-sulfatase |
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| YU Yong-li et al.Cloning and Expression of Flavobacterium heparinum 2-O-sulfatase[J].Journal of Anhui Agricultural Sciences,2011,39(12):7286-7288. |
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| Authors: | YU Yong-li |
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| Institution: | YU Yong-li et al(School of Chemical Engineering,Wuhan University of Technology,Wuhan,Hubei 430070) |
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| Abstract: | Objective] To clone and express 2-O-sulfatase gene from Flavobacterium heparinum.Method] The 2-O-sulfatase gene was amplified with PCR from the genomic DNA of Flavobacterium heparinum,and it was linked into expression plasmid pET-28a after sequencing,then the recombined plasmid was expressed in E.coli BL21(DE3).Result] The recombined pET-28a(+)with the PCR product of 2-O-sulfatase which was confirmed by DNA sequencing was successfully constructed.The expression product of the recombined plasmid pET-28a i... |
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| Keywords: | 2-O-sulfatase |
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