小鼠SRS白血病病毒cDNA基因库的构建  

CONSTRUCTION OF cDNA LIBRARIES OF SRS-MLV GENOME

作　　者：季文琴[1] 付四清[1] 程立[1] 黄春妹 张萌[2] 宋后燕[2] 

机构地区：[1]上海医科大学基础医学部病理生理学教研室 [2]上海医科大学分子遗传学教研室

出　　处：《上海医科大学学报》1990年第1期15-18,共4页Journal of Fudan University(Medical Science)

基　　金：国家自然科学基金

摘　　要：从小鼠SRS腹水上清液中分离出小鼠SRS白血病病毒基因组RNA,以oligo(dT)_(18)为引物,在逆向转录酶和DNA聚合酶I催化下合成cDNA,经S1核酸酶处理及3'-OH端多聚dC化后,与3'-OH端多聚dG化载体DNA形成重组DNA分子,转化相应的受体菌,构建成小鼠SRS白血病病毒cDNA基因库。结果表明小鼠SRS白血病病毒基因组RNA含有polyA^+结构,与Moloney小鼠白血病病毒基因组有一定的同源性,并在SRS腹水瘤细胞中有转录活性。提示SRS白血病病毒与SRS瘤细胞的发生和发展有关。SRS-MLV genomio RNA was separated and purified from the aScitio supernatant of SRS mice. Primed by oligo(dT), the viral genomio RNA were reverse-transoripted into oDNA with reverse transoriptase and DNA polymerase I. Poly (dO) were added to the 3'-hydroxyl termini of the ds-cDNA which had been treated with S-l nucleate and a corresponding number of poly(dG) were added to the 3'-hydroxyl termini of the linearized vector DNA. The tailed vector DNA and the tailed cDNA annealed and transformed E. coli cells. Thus we had constructed SRS-MLV genomio oDNA libraries and screenedfor several positive oDNA clones. We had also identified that SRS-MLV genomio RNA contained poly A structure and was homologous to Moloney-MLV genome. In SRS asoitio sarcoma cells was a higher transcriptive level of SRS-MLV genome, it was suggested that SRS-MLV was involved in the initiation and development of SRS ascitio sarcoma cells.

关 键 词：白血病病毒 CDNA 基因库 

分 类 号：Q784[生物学—分子生物学]