可溶性E-选择素cDNA基因克隆及其真核表达  

Cloning and expression of soluble E selectin cDNA in vaccinia virus system

作　　者：胡美茹[1] 蔡铀庆[1] 汲言山[1] 孙涛[1] 王建安[1] 孙英勋[1] 沈倍奋[1] 

机构地区：[1]军事医学科学院基础医学研究所

出　　处：《军事医学科学院院刊》1996年第3期182-184,共3页Bulletin of the Academy of Military Medical Sciences

基　　金：国家863青年基金

摘　　要：从ｐＵＣ１８／Ｅ－选择素重组质粒经ＰＣＲ扩增得到可溶性Ｅ－选择素ｃＤＮＡ基因，将此基因插入到痘苗病毒表达载体ｐＪＳＡ１１７５的ＳｍａⅠ位点，构建了正向表达质粒ｐＪＳＡ１１７５／可溶性Ｅ－选择素，采用脂质体共转染的方法，将上述质粒转染ＴＫ－１４３细胞，用ＢｕｄＲ和Ｘ－ｇａｌ双筛选获得带有人可溶性Ｅ－选择素ｃＤＮＡ重组痘苗病毒，连续传代１０代以上，经ＡＰＡＡＰ检测和ＳＤＳ－ＰＡＧＥ电泳，证实重组痘苗病毒能稳定。Soluble E selectin is an important modulator in adhesion and an indicator in diseases such as infection, autoimmune diseases and tumor metastasis. Soluble E selectin cDNA was amplified by PCR from the plasmid pUC18/E selectin constructed by this laboratory, and which was then inserted at SmaⅠ site of the plasmid pJSA1175 in positive orientation. TK - 143 cell line was transfected with the plasmid pJSA1175/soluble E selectin and wild vaccinia virus by lipofectin cotransfection method and the recombinant vaccinia virus was screened with BudR and X gal, purified and identified by PCR. The recombinant vaccina virus obtained can express soluble E selectin stably as demonstrated by APAAP and SDS PAGE.

关 键 词：可溶性 E-选择素 基因 痘病毒 感染 真核细胞 

分 类 号：R394.8[医药卫生—医学遗传学] R373.12[医药卫生—基础医学]