Cloning of human sperm protein(SP17) and expression in escherichia coli DH5α |
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Authors: | WANG Min-zhen LIU Zhi-cheng GAO Zhan ZHENG Wen-bo LUO Jian-hong |
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Institution: | 1. Sir Run Run Shaw Hospitol,Medical Academy of Zhejiang University, Hangzhou 310031, China;
2. Molecular Biology Laboratory, Medical Academy of Zhejiang University, Hangzhou 310031, China |
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Abstract: | AIM: To obtain GST fusion protein of hSP17 gene and construct the recombinant plasmidfor expression in E.coli.METHODS:Total fragment of hS P17 cDNA gene were amplified by RT-PCR,then subcoloned into p GEX-3b to generate recombinant hS P17/pGEX.Right orientation of insert are identified by restricted enzyme digestion.Transform the correct recombinant plasmid into the E.coli DH5a.The expression of fusion proteins hS P17-GST were induced by adding isopropylthiogalactoside(IPTG).RESUL TS and CONCLUSION:The recombinant plasmid hS P17/pGEX-3b could express effectively in E.coli and a high level of fusion protein hsp17-GST with the predicted molecular weight was detected. |
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Keywords: | Human Sperm Proteins Genes |
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