人T细胞活化衔接因子基因真核表达载体的构建  

Construction of human LAT gene eukaryotic expression vector

作　　者：任涟萍[1] 郭雪君[1] 

机构地区：[1]上海交通大学医学院附属新华医院呼吸内科,200092

出　　处：《国际免疫学杂志》2009年第5期337-341,共5页International Journal of Immunology

基　　金：上海市科委基础研究重点项目（04JC14048）

摘　　要：目的构建人T细胞活化衔接因子（LAT）基因的真核表达载体。方法以人外周血单个核细胞的cDNA为模板，采用聚合酶链式反应（PCR）扩增LAT基因编码区的全部序列，克隆人真核细胞表达载体pCMV—Myc中，测序鉴定目的基因并运用核转染的方法转染人外周血T淋巴细胞。结果PCR扩增的特异性片段长度为729bp，以此构建重组质粒pCMV—Myc—LAT，测序结果与Genbank中的人LAT基因cDNA序列一致。转染人外周血T淋巴细胞后可检测到LAT蛋白的表达。结论成功构建了pCMV—Myc—LAT重组真核表达载体。To construct the eukaryotic expression vector of human linker for activation of T cells （LAT） gene. Methods LAT eDNA of human peripheral blood mononuclear cells was amplified with polymerase chain reaction and inserted into the eukaryotic expression vector of pCMV-Myc. The recombinant plasmid was verified by DNA sequencing and used to transfeet human T cells. Results The length of ampli- fied fragment was 729 bp. The sequence of LAT gene was confirmed by Genbank. The LAT protein was detected in human T cells. Conclusion The recombinant eukaryotic expression vector pCMV-Myc-LAT was successfully constructed.

关 键 词：LAT 基因克隆 真核表达载体 

分 类 号：Q78[生物学—分子生物学]