2004Harbin Yike Daxue xuebaoRequires access

Expression,purification and identification of recombinant GST-p16 fusion protein

Cong Shi

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Abstract

Objective To express,purify and identify recombinant GST p16 fusion protein using GST gene fusion system.Methods The cDNA of p16 was amplified from plasmid pM p16 and cloned into pGEX 6P 1 expression vector.The recombinant plasmid pGEX p16 was expressed in E.coli BL21 cells and the products were purified by GST purifying system.The specific expression was identified by Western blot.Results The resolved GST p16 fusion protein on 15% SDS PAGE showed a major band at position of 42kD and the fusion protein was recognized by anti p16 antibody on PVDF membrane.Conclusion The recombinant GST p16 fusion protein is expressed in E.coli cells efficiently,and the purified GST p16 fusion protein can be used in further study.

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Objective To express,purify and identify recombinant GST p16 fusion protein using GST gene fusion system.Methods The cDNA of p16 was amplified from plasmid pM p16 and cloned into pGEX 6P 1 expression vector.The recombinant plasmid pGEX p16 was expressed in E.coli BL21 cells and the products were purified by GST purifying system.The specific expression was identified by Western blot.Results The resolved GST p16 fusion protein on 15% SDS PAGE showed a major band at position of 42kD and the fusion protein was recognized by anti p16 antibody on PVDF membrane.Conclusion The recombinant GST p16 fusion protein is expressed in E.coli cells efficiently,and the purified GST p16 fusion protein can be used in further study.

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Available abstract

Objective To express,purify and identify recombinant GST p16 fusion protein using GST gene fusion system.Methods The cDNA of p16 was amplified from plasmid pM p16 and cloned into pGEX 6P 1 expression vector.The recombinant plasmid pGEX p16 was expressed in E.coli BL21 cells and the products were purified by GST purifying system.The specific expression was identified by Western blot.Results The resolved GST p16 fusion protein on 15% SDS PAGE showed a major band at position of 42kD and the fusion protein was recognized by anti p16 antibody on PVDF membrane.Conclusion The recombinant GST p16 fusion protein is expressed in E.coli cells efficiently,and the purified GST p16 fusion protein can be used in further study.

Key concepts: Fusion protein, Recombinant DNA, Molecular biology, Myc-tag, Western blot, Complementary DNA, Plasmid, Fusion gene

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