杀菌_渗透增加蛋白；基因表达；大肠杆菌_____\]

人杀菌/渗透增强蛋白N末端片段表达载体的构建

http://www.100md.com 《郧阳医学院学报》 2006年第3期

杀菌,渗透增加蛋白；基因表达；大肠杆菌,,\],杀菌,渗透增加蛋白；基因表达；大肠杆菌,1材料和方法,2结果,3讨论,[参考文献\]

     [摘要\]目的:建立杀菌/渗透增强蛋白(BPI) N端193个氨基酸的重组表达质粒。方法:利用RTPCR的方法从HL60细胞内扩增出BPI氨基端1～193个氨基酸的基因序列，克隆入T载体。将BPI 193基因片段定向克隆入原核表达载体pET28a中，构建重组的原核表达质粒pETBPI 193，转化大肠杆菌BL 21菌株。结果:从HL60细胞中扩增得到579 bp的BPI 193基因片段，构建了TBPI 193亚克隆和PETBPI 193重组表达质粒。结论:成功构建了pETBPI 193重组表达质粒。

    [关键词\] 杀菌/渗透增加蛋白；基因表达；大肠杆菌

    The Construction of Human Bactericidal/Permeability Increasing Protein Nterminal Fragment Expression Vector

    DONG Wei, ZHAN Zhen, TONG Shu-juan.(Nanjing University of Traditional Chinese Medicine，Nanjing,Jiangsu 210046,China)

    Abstract: Objective To construct a prokaryotic expression vector encoding first 193 amino acids of BPI Nterminal. Methods Total RNA was extracted from HL60 and then was amplified by using RTPCR. The PCR product was cloned into pMD18T vector. BPI 193 cDNA was directly inserted into pET28a plasmid with T4 DNA ligase. pETBPI 193 recombinant expression vector was transformed into competent E.coli BL 21 and protein expression was induced by IPTG. Results A 579 bp fragment was amplified by RTPCR. BPI cDNA was inserted into pET28a.Conclusions pETBPI 193 recombinant expression vector was constructed successfully. ......

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