p16基因的原核表达及抗P16抗血清的制备  

Expression of p16 as fusion protein in E. coli and production of antisera to GST-P16

作　　者：覃林花[1] 黄长晖[1] 博继梁 

机构地区：[1]第二军医大学医学分子遗传学军队重点实验室

出　　处：《生物技术通讯》1998年第3期177-179,共3页Letters in Biotechnology

基　　金：国家自然科学基金(项目号 39670400)

摘　　要：采用基因重租技术,将野生型p16 cDNA通过中间载体pVL1392最后载入表达载体pGEX-5T,IPTG诱导重组质粒转化的大肠杆菌,蛋白质印迹证实42×10~3的融合蛋白GST-P16的表达。表达了GST-P16的重组菌经加热破菌后行SDS-PAGE,将GST-P16蛋白条带切下后经反复冻融于皮内多点注射免疫家兔。所收获的兔血清经蛋白质印迹法检测到抗P16抗体滴度为1:625。结果表明通过pGEX-5T融合蛋白表达系统能方便地提供制备抗P16抗血清的免疫原,该免疫原未经纯化并未影响抗P16抗血清的产生。p16 cDNA was cloned into expression vector pGEX-5T and transformed E. coli TG1. After 3 hours induced by IPTG, the expression of recombinant 42 kD GST-pl6 was confirmed by SDS-PAGE and Western blotting. For producing antibody to p16. the recombinant GST-p16 fusion protern belt was cut out from SDS-PAGE gel and emulsified with adjuvant to be as antigen to inoculate rabbits. After the rabbits were subcutaneous injected with antigen protein about 0. 5 mg in 0 day, 28 day and 56 day, the titer of anti-p16 antisera reached to 1 : 625 when examined by Western blotting.

关 键 词：p16基因(CDKN2) 原核表达 蛋白免疫 

分 类 号：R346[医药卫生—基础医学]