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cory0931

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巫山云

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注册: 2012-02-29
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专业: 临床兽医学

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爱与雨下: 金币+1 2013-03-19 21:08:24
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phu_grassman: 金币+1, 翻译EPI+1, thanks for your help/ 2013-04-04 09:21:21

小试一下，仅供参考；
To obtain the fusion protein of swine interleukin-6 & interferon-α(IL6 & IFNα) and explore its feasibility of using as an immunoadjuvant, primers with restriction endonuclease sites, linker and his-tag was designed according to the sequences of Il6/IFNα and vector pET32a. Matured peptide sequences of swine IL6 & IFNαwere successfully cloned and amplified by transfecting into E. coli through pMD18, respectively. The pMD18 plasmids were extracted and enzymatically cut to build the recombinant plasmid pMD18-IL6-IFNα(rpMD18-IL6-IFNα), followed by transfection into competent E. coli DH5α and E. coli Rosetta(DE3), and expressed by different IPTG concentration induction in different time. The results showed that the fusion expression plasmids were successfully built and highly expressed in E. coli Rosetta(DE3) according to SDS-PAGE detection. Molecular weight of the interest protein is 44.96kDa, which is agree with predicted value, and is mainly existed as insoluble inclusion bodies by SDS-PAGE detection after ultrasound treatment.

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