摘要
目的研究干扰素诱导蛋白IFIT1的原核可溶性表达。方法通过逆转录聚合酶链反应(RT-PCR)获得IFIT1编码序列,克隆入原核表达载体,电穿孔转化大肠杆菌,筛选阳性克隆,诱导细菌表达,蛋白凝胶电泳观察。结果构建的原核表达平台实现了较丰富的麦芽糖结合蛋白融合IFIT1(MBP-IFIT1)可溶性表达。结论 MBP-IFIT1的原核可溶性表达为后续IFIT1蛋白结合分析及免疫分析提供了材料及方法学基础。
Objective To investigate the soluble prokaryotic expession of interferon induced factor with tetratricopeptide repeats-1.Methods By reverse transcription PCR,murine IFIT1 cDNA coding sequence was acquired,and cloned into the prokaryotic expression vector.Then the E.coli was transformed by electroporation with the recombinant.The screened positive cloning was induced to express target recombining protein which was observed with SDS-PAGE.Results The aplenty soluble fusion proteins MBP-IFIT1 could be obtained by this prokaryotic expression construction.Conclusion Accomplishment of the soluble expression of MBP-IFIT1 provides basements of the IFIT1 follow-up functional studies.
出处
《重庆医学》
CAS
CSCD
北大核心
2011年第19期1873-1875,共3页
Chongqing medicine
基金
国家973创伤项目05课题基金资助项目(2005CB522605)
海外青年学者合作研究基金资助项目(30328025)
关键词
创伤和损伤
细胞应激
重组
原核表达
wounds and injuries
cell-stress
recombinants
prokaryotic expression
