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《Progress in Veterinary Medicine》 2018-06
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Prokaryotic Expression and Analysis of Truncated IBV S1 Gene

WANG Shan-shan;LI Shu-dong;LIU Huan-huan;GAO Guo-qiang;HOU Xi-lin;College of Animal Science and Technology,Heilongjiang Bayi Agricultural University,Animal Infectious Disease Diagnosis and New Vaccine Laboratory;  
The target fragment of 468 bp of IBV H120 S1 gene was amplified by RT-PCR.The fragment was cloned into pQE-30 prokaryotic expression vector and the recombinant plasmids were transformed into competent BL21(DE3)Escherichia coli,followed by recombinant protein expression using IPTG induction.To obtain the purified recombinant protein,the concentration of IPTG and the induction time were optimized.The Western blot was utilized to verify the activity of the protein.The results showed that pQE-30-S1 recombinant strain was successfully constructed,and the target protein with the size of about 18 ku was expressed by IPTG induction.The optimal induction concentration was 0.2 mmol/L and the optimum induction time was 4 h.The protein mainly existed in inclusion bodies,and Western blot showed that the recombinant protein could react specifically with IBV polyclonal antibodies.
【Fund】: 研究生创新项目(YJSCX2016-Y25)
【CateGory Index】: S852.65
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