小鼠分泌型γ干扰素原核表达载体pFLAG-IFN-γ的构建

Construction of prokaryotic expression vector pFLAG-IFN-γ with mouse secretion type IFN-γ

目的构建小鼠分泌型γ干扰素(IFN-γ)原核表达载体pFLAG-IFN-γ。方法利用分子生物学方法,从pcD-NA3.1-Egr1-IFN-γ质粒上将IFN-γ切下,然后连接到原核表达载体pFLAG-shift12TM上,构建成pFLAG-IFN-γ表达载体,进行PCR、酶切鉴定。结果经鉴定,所构建的质粒pFLAG-IFN-γ的鉴定结果与预期的一致。结论成功地构建了带有分泌信号的原核表达载体pFLAG-IFN-γ。

Objective To construct the prokaryotic expression vector pFLAG-IFN-γ which contains the mouse secreting type interferon-gamma （IFN-γ） .Methods The IFN-γ fragment was acquired from the plasmid pcDNA3.1-Egrl-IFN-γ,and then it was ligated to the prokaryotic expression vector pFLAG-shift12^TM which finally constructed the recombinant plasmid pFLAG-IFN-γ）. Further-more, PCR and enzyme digestion were used to identify it. Results The fragments acquired by PCR and enzyme digestion identification were all in concordance with the expected results that indicated that the pFLAG-IFN-γ was correctly constructed. Conclusion The recombinant plasmid pFLAG-IFN-γ which contains mouse secretion type IFN-γ was constructed successfully.