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人lrpcDNA全长编码区的克隆及其在大肠杆菌中的表达
http://www.100md.com 《第四军医大学学报》 2006年第5期
人lrp,,脂多糖;,应答基因;逆转录聚合酶链反应;基因表达,0引言,1材料和方法,2结果,3讨论,【参考文献】
     length coding region of human lipopolysaccharide responsed gene cDNA and its expression in E.coli

    SONG QingHe, CHAI YuBo, CHEN SuMin, CHEN NanChun, HUANG Yong, DAI ZhongMing

    Department of Biochemistry and Molecular Biology, School of Basic Medicine, Fourth MilitaryMedical University, Xian 710032, China

    【Abstract】 AIM: To clone, express and identify fulllength human lipopolysaccharide (LPS) responsed gene (lrp)cDNA coding sequence. METHODS: Total RNA was extracted from LPSstimulated human embryonic kidney cells (HEK293) and the fulllength human lrpcDNA sequence was obtained by RTPCR. The lrpcDNA coding sequence was cloned into pcTAT fusion expression vector, then transferred into E.coli BL21 and induced to express with IPTG. The expressed fusion protein (6HisTATLrp) was analyzed by SDSPAGE and Western blot. RESULTS: DNA sequencing result showed that the lrpcDNA coding sequence we cloned was not exactly consistent with that issued by GenBank. SDSPAGE analysis demonstrated that the 6HisTATLrp fusion protein was expressed successfully in E.coli. The fused protein band amounted to 17% of the total bacteria protein and the expressed protein reacted with antisera. CONCLUSION: Human lrpcDNA fulllength coding sequence is successfully cloned and expressed, which offers a basis for further research of lrp function. ......

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