抑瘤素McDNA的克隆及表达研究  

Study on Molecular Cloning and Primary Expression of Oncostatin M

作　　者：段海清[1] 张兆山[1] 董自正[1] 曹勇[1] 黄翠芬[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《生物化学杂志》1997年第3期259-263,共5页

摘　　要：抑瘤素M（onecostatinM，OSM）是一种多功能的细胞生长调节因子．从PMA刺激后的U937细胞系提取总RNA，采用逆转录-PCR方法分离到了抑瘤素M的cDNA；将抑瘤素M的cDNA克隆到质粒pUC19中，筛选三个阳性克隆进行序列分析，与国外报导序列完全一致；将抑癌素M的cDNA克隆到质粒pBV220后再转化DH5a进行模拟表达，SDS-PAGE分析表明有OSM表达，表达量约占细菌总蛋白5％，经过初步纯化的OSM能明显抑制A375细胞的生长．Oncostatin M (OSM) is a polypeptide cytokine having diverse biological activity. OSM cDNA was isolated from total RNA of U937 cells treated with PMA through reverse transcription and polymerase chain reaction (RT-PCR). Sequence analysis of four positive coines shows that there are three clones having the same sequence with the reported previlusly and the other one has a point mutation at the 46th threonine codon (ACA to ACG). Cloning the cDNA encoding sequence (without the signal peptide sequence) into plasmid pBV220 and transformating DH5a resulted expression level of the rhOSM was about 5% of the total cell protein by SDSPAGE analysis. The crude rhOSM can inhibit the growth of A 375 cells greatly.

关 键 词：抑瘤素M 基因表达 CDNA 基因克隆 

分 类 号：Q523.03[生物学—生物化学] R384.8[医药卫生—医学寄生虫学]