| 大肠杆菌二硫键异构酶DsbA和脯氨酸异构酶PPIaseA双顺反子表达载体的构建及其在大肠杆菌中的表达 |
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| 引用本文： | 华子春,许立,孙爱龙,金冬生.大肠杆菌二硫键异构酶DsbA和脯氨酸异构酶PPIaseA双顺反子表达载体的构建及其在大肠杆菌中的表达[J].南京大学学报(自然科学版),1998(2). |
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| 作者姓名： | 华子春 许立 孙爱龙 金冬生 |
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| 作者单位： | 南京大学生物化学系，江苏省中医药研究所 |
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| 摘 要： | 用ＰＣＲ方法获得大肠杆菌二硫键异构酶ＤｓｂＡ的编码基因ｄｓｂＡ和大肠杆菌脯氨酸异构酶ＰＰＩａｓｅＡ的编码基因ｒｏｔ，并将ＤｓｂＡ和ｒｏｔ以双顺反子形式克隆至含有Ｐｔａｃ启动子的表达载体ｐＫＫ２３３－２中。在ＩＰＴＧ的诱导下，ＤｓｂＡ和ＰＰＩａｓｅＡ获得了表达。ＳＤＳ－ＰＡＧＥ和薄层扫描分析表明：ＤｓｂＡ、ＰＰＩａｓｅＡ的表达水平分别为占菌体裂解上清液总蛋白质的４．３２％和４．０６％。
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| 关 键 词： | 脯氨酸异构酶，二硫键异构酶，双顺反子 |
Construction of a dicistronic expression plasmid expressing E.coli protein disulfide isomerase dsba and E.coli peptidylprolyl cis,transisomerase ppiasea and its expression in E.coli |
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| Hua Zichun,Xu Li,Sun Ailong,Jin Dongsheng.Construction of a dicistronic expression plasmid expressing E.coli protein disulfide isomerase dsba and E.coli peptidylprolyl cis,transisomerase ppiasea and its expression in E.coli[J].Journal of Nanjing University: Nat Sci Ed,1998(2). |
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| Authors: | Hua Zichun Xu Li Sun Ailong Jin Dongsheng |
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| Institution: | Hua Zichun 1) Xu Li 2) Sun Ailong 1) Jin Dongsheng 2) |
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| Abstract: | The genes dsbA,encoding E.coli protein disulide isomerase DsbA,and rot,encoding E.coli peptidylprolyl cis,trans isomerase were obtained by PCR method and cloned in dicistronic gene organization into plasmid pKK233 2 containing Ptac promoter.Upon IPTG induciton,DsbA and PPIaseA were expressed in E.coli.SDS PAGE analysis and densitometric sccanning showed that the expression levels of DsbA and PPIaseA were 4.32% and 4.06% of total cellular lysate proteins respectively. |
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| Keywords: | protein disulfide isomerase peptidylprolyl cis trans isomerase dicistron |
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