线虫fat-1基因的原核表达、纯化及其抗体的制备  

Prokaryotic expression of the C.elegans fat-1 gene and preparation of antibody

作　　者：张鹏[1] 曹立雪[2] 陈宏[1] 马润林[2] 

机构地区：[1]西北农林科技大学动物科技学院陕西省农业分子生物学重点实验室,陕西杨凌712100 [2]中国科学院遗传与发育生物学研究所,北京100101

出　　处：《西北农林科技大学学报（自然科学版）》2007年第1期24-28,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：中国科学院知识创新工程青年科学家专项基金

摘　　要：通过RT-RCR得到线虫f a t-1基因全长,将其N端亲水区克隆至原核表达载体pGEX-4T-2中,构建重组质粒pGEX-f a t1-N,将其转化大肠杆菌,并进行诱导表达。对表达产物G ST-f a t1-N融合蛋白进行纯化,制备其抗体,并对抗体效价进行了检测。结果表明,线虫f a t-1基因能在大肠杆菌中表达,制备的抗体能识别在原核表达系统内表达的G ST-f a t1-N融合蛋白,且效价很高,达到1∶107。Caenorhabditis elegans fat 1 gene was obtained by RT-PCR,and the sequences of fat-1 gene N-terminal were cloned into fusion expression vector pGEX-4T-2 to generate recombinant plasmid pGEX- fat1-N. After expressing in Escherichia coli,the expressed product GST-fat1-N fusion protein was purified and used to immunize rabbit. The result showed C. elegans fat-1 gene could be expressed in E. coli,and the antibody against GST-fat1-N protein had a good affinity and selectivity by enzyme linked immunosorbent assay.

关 键 词：fat-1基因 线虫 原核表达 抗体制备 

分 类 号：Q786[生物学—分子生物学]