中国免疫学杂志Issue(10)：1370-1374,5.DOI:10.3969/j.issn.1000-484X.2015.10.016
GST－NRP－1融合蛋白的原核表达及纯化
Prokaryotic expression and purification of GST-NRP-1 fusion protein
摘要
Abstract
Objective:To construct GST-tagged human NRP-1 fusion protein expression vector and induce its expression in Escherichia coli ( E.coli) ,then carry on inclusion body refolding and purification so as to obtain GST-NRP-1 fusion protein.Methods:NRP-1 gene was amplified by RT-PCR and inserted into pCR-blunt vector.Then the reconstructed plasmid was inserted into prokaryotic expression vector pGEX-4T-1.The constructed pGEX-4T-1-NRP-1 expression vector was transformed into BL21 cells and induced by i-sopropyl-β-D-thiogalactoside ( IPTG).Bacterial bodies were disrupted by sonication.Then the soluble fraction of fusion proteins were verified by Western blot and purified by Glutathione Sepharose 4B after inclusion body refolding.Results: The NRP-1 gene fragment was amplified by RT-PCR and inserted into pCR-blunt vector.Fusion protein expression vector pGEX-4T-1-NRP-1 was constructed suc-cessfully.After transformation, GST-NRP-1 expression vector was detected in BL21 cells and obtained purifying protein after refolding.Conclusion:The plasmid GST-NRP-1 was constructed successfully and laid basis for subsequent studies.关键词
NRP-1/原核表达/融合蛋白/纯化Key words
NRP-1/Prokaryotic expression/Fusion protein/Purification分类
医药卫生引用本文复制引用
韩正祥,张梦瑾,徐杰,王红梅,杜秀平,陈翀,徐开林..GST－NRP－1融合蛋白的原核表达及纯化[J].中国免疫学杂志,2015,(10):1370-1374,5.基金项目
本文受国家自然科学基金（ No．30901753）和江苏省“六大人才高峰”B类项目资助（No．2014－WSW－040）。 （）
