小鼠SRS白血病病毒cDNA基因库的构建

CONSTRUCTION OF cDNA LIBRARIES OF SRS-MLV GENOME

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.