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《China Dairy Cattle》 2009-01
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Construction of Procaryotic Expression Vector of Bactericidal/permeability-increasing Protein N-terminal Gene of Holstein Cow

Gao Heng1, Song Yanhua1, Wen Naxiang1, Qi Kezong2 (1.Guangdong Wen’s Group, Xinxing 527439;2.College of Animal Science & Techology, Anhui Agricultural University, Hefei, 230036)  
According to the method provided by Gao Heng[5],the N-terminal gene was amplified by RT-PCR from mRNA which were extracted from the polymorphonuclear neutrophils of holstein cow. The purified DNA was connected with expression vector pGEX-4T-1, constructed recombinant plasmid pGEX-4T-1-BPI, which was transformed into E.Coli BL21. It could produce fusion protein induced with IPTG. The results showed that A 714 bp fragment was acquired, and there was a basic mutated group in 503rd compared with the reports. The E.coli with recombinant plasmid was induced with IPTG. The products were analyzed by SDS-PAGE, and a new protein of 52kD was detected. Its molecular weight was the same as expected. In conclusion, the purified recombinant BPI was obtained successfully in our experiment.
【Fund】: 国家“863”计划(2006AA10Z320);; 安徽省教育厅自然科学研究项目(KJ2007A021)
【CateGory Index】: S858.23
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