| High-level expression of whiG——A key gene for Streptomyces differentiation in Escherichia coli |
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| 引用本文： | 谭华荣,田宇清,杨海花,吴畏,董可宁,K. F. Chater and M. J. Buttner. High-level expression of whiG——A key gene for Streptomyces differentiation in Escherichia coli[J]. 中国科学：生命科学英文版, 1996, 39(3): 284-290 |
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| 作者姓名： | 谭华荣 田宇清 杨海花 吴畏 董可宁 K. F. Chater and M. J. Buttner |
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| 作者单位： | Institute of Microbiology，Chinese Academy of Sciences，Beijing 100080，China，Institute of Microbiology，Chinese Academy of Sciences，Beijing 100080，China，Institute of Microbiology，Chinese Academy of Sciences，Beijing 100080，China，Institute of Microbiology，Chinese Academy of Sciences，Beijing 100080，China，Institute of Microbiology，Chinese Academy of Sciences，Beijing 100080，China，John Innes Institute，Norwich，England) |
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| 基金项目： | Project supported by the National Natural Science Foundation of China and the Foundation of the Royal Society of UK. |
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| 摘 要： | Six nucleotides located in the region of translation start site of whiG were changed. whiG was amplified by PCR technique. Reformed sequences were determined. This gene was directly subcloned into expression vector pET11c containing strong T7 promoter, and the recombinant plasmid was introduced into E. coli BL21(DE3), which could be induced by IPTG to produce T7 RNA polymerase. The SDS-PAGE result showed that whiG highly expressed in E. coli BL21(DE3), and the yield of whiG product was about 20% of insoluble proteins in cell. whiG product (σwhiG) was further identified by Western blot hybridization after making its antibody. whiG gene was subcloned into Streptomyces plasmid pIJ6021, and then it was introduced into sporulation deficient mutant C71 from Streptomyces coelicolor. The result showed that C71 could restore sporulation and σwhiG has biological functions.
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| 关 键 词： | Streptomyces differentiation σ~(whiG) high expression. |
| 收稿时间： | 1995-12-01 |
High-level expression of whiG--A key gene for Streptomyces differentiation in Escherichia coli |
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| TAN Huarong TIAN Yuqing YANG HaihuaWU Wei DONG KeningK. F. Chater M. J. Buttner. High-level expression of whiG--A key gene for Streptomyces differentiation in Escherichia coli[J]. Science China Life sciences, 1996, 39(3): 284-290 |
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| Authors: | TAN Huarong TIAN Yuqing YANG HaihuaWU Wei DONG KeningK. F. Chater M. J. Buttner |
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| Affiliation: | 1.Institute of Microbiology; Chinese Academy of Sciences; Beijing 100080; China2.John Innes Institute; Norwich; England |
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| Abstract: | Six nucleotides located in the region of translation start site of whiG were changed. whiG was amplified by PCR technique. Reformed sequences were determined. This gene was directly subcloned into expression vector pET11c containing strong T7 promoter, and the recombinant plasmid was introduced into E. coli BL21(DE3), which could be induced by IPTG to produce T7 RNA polymerase. The SDS-PAGE result showed that whiG highly expressed in E. coli BL21(DE3), and the yield of whiG product was about 20% of insoluble proteins in cell. whiG product (σwhiG) was further identified by Western blot hybridization after making its antibody. whiG gene was subcloned into Streptomyces plasmid pIJ6021, and then it was introduced into sporulation deficient mutant C71 from Streptomyces coelicolor. The result showed that C71 could restore sporulation and σwhiG has biological functions. |
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| Keywords: | Streptomyces differentiation &sigma whiG high expression. |
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