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始旋链霉菌HCCB2003-8 snaA、snaB和snaC基因的克隆、序列分析及原核表达
引用本文:王宁波,殷瑜,黄为一,朱丽,陈代杰. 始旋链霉菌HCCB2003-8 snaA、snaB和snaC基因的克隆、序列分析及原核表达[J]. 中国抗生素杂志, 2006, 31(11): 654-656,674
作者姓名:王宁波  殷瑜  黄为一  朱丽  陈代杰
作者单位:1. 南京农业大学微生物学系,南京,210095
2. 上海来益生物药物研究开发中心,上海,201203
3. 上海来益生物药物研究开发中心,上海,201203;上海医药工业研究院,上海,200040
摘    要:以始旋链霉菌(Streptomyces pristinaespiralis)HCCB2003—8的染色体DNA为模板,通过优化PCR条件扩增出FMN还原酶、PII.合酶的基因片段。与文献报道的基因序列做同源比较,snaA和snaB碱基序列分别只有一个碱基的差异,snaC碱基序列完全匹配。将snaA、snaB和snaC的基因片段插入原核表达载体pET30a中,并转化至大肠埃希菌BL21(DE3)。SDS—PAGE结果表明。经IPTG诱导后基因分别得到表达。

关 键 词:始旋链霉菌  普那霉素  基因克隆  表达
文章编号:1001-8689(2006)11-0654-03
收稿时间:2006-05-15
修稿时间:2006-05-22

Cloning, sequencing and prokaryotic expression of snaA, snaB and snaC gene from Streptomyces pristinaespiralis HCCB2003-8
Wang Ning-bo,Ying Yu,Huang Wei-yi,Zhu Li,Chen Dai-jie. Cloning, sequencing and prokaryotic expression of snaA, snaB and snaC gene from Streptomyces pristinaespiralis HCCB2003-8[J]. Chinese Journal of Antibiotics, 2006, 31(11): 654-656,674
Authors:Wang Ning-bo  Ying Yu  Huang Wei-yi  Zhu Li  Chen Dai-jie
Abstract:Through PCR method and optimization of PCR reaction system,snaA,snaB and snaC gene were obtained from Streptomyces pristinaespiralis HCCB2003-8.Analysis of snaA,snaB and snaC gene(sequences) showed that there was only one base,different from the reported sequences of snaA and snaB respectively.snaC sequence was identical to the reported sequence.snaA,snaB and snaC gene were inserted into pET30a vector and were transformed into E.coli BL21(DE3) strain.After induced by IPTG,the result of SDS-PAGE showed that they were expressed.
Keywords:Streptomyces pristinaespiralis   Pristinamycin   Gene cloning   Expression
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