南方医科大学学报2011，Vol.31Issue(7)：1232-1235,4.
人CD96基因胞外区的原核表达及抗血清制备
Human CD96 gene cloning, expression and identification
摘要
Abstract
Objective To construct and express human CD96 gene outer membrane domain (hCD96om) in prokaryotic cells and prepare rabbit polyclonal antibody of hCD96om. Methods hCD96om was amplified by RT-PCR from the peripheral blood of patients with acute myeloid leukemia and inserted into prokaryotic expression vector pET32a(+) to construct the recombinant plasmid pET32-CD96. The expression of hCD96om was induced by IPTG in BL21(DE3) cells, and the expression product was identified by Western blotting. The anti-hCD96 polyclonal antibody was prepared by immunization of rabbits with the fusion protein. The specificity of anti-hCD96 antibody was determined by Western blotting. Results hCD96om protein was expressed in E.coli BL21(DE3) cells in the form of inclusion body, with a relative molecular mass around 37 kD. Western blotting showed a specific reaction of the prepared antiserum with the 70 kD protein extracted from human leukemia cell line HL-60 cells and with the 37 000 hCD96om fusion protein. Conclusion The CD96 gene of human has been successfully cloned and expressed in BL2K1(E3) cells, and its rabbit polyclonal antibody has been obtained.关键词
人CD96基因/原核表达/白血病干细胞/多克隆抗体Key words
human CD96 gene/prokaryotic expression/leukemic stem cell/polyclonal antibodies分类
医药卫生引用本文复制引用
曾建明,陈茶,刘飞,谭坪海,王丽娜,李沫,陈中华,李松,龙一飞,李有强..人CD96基因胞外区的原核表达及抗血清制备[J].南方医科大学学报,2011,31(7):1232-1235,4.基金项目
广东省医学科学技术研究基金(B2008066) （B2008066）
