Legrand A, Nachtigal M
Department of Pathology, University of South Carolina, School of Medicine, Columbia 29208.
Intervirology. 1991;32(3):173-84. doi: 10.1159/000150197.
The plasmid pSV3-neo that carries the SV40 early genetic region and the neor resistance gene underwent homologous recombination between SV40-duplicated sequences of the small t antigen intron and the polyadenylation site after transfection into rabbit vascular smooth muscle cells. This recombination yielded two free plasmids of 1.7 and 3.0 kb that replicated in transfected rabbit cells, while the vector plasmid did not. Replication of the two recombinant plasmids continued for several months in rabbit cells transformed by pSV3-neo, and their structure remained unchanged. Between 15 and 2,510 copies per cell of the 3.0-kb plasmid and between 370 and 1,565 copies per cell of the 1.7-kb plasmid were found in different pSV3-neo-transformed rabbit clones. Similar recombined plasmids were found 3 days after transfection of pSV3-neo onto SV40-transformed rabbit cells. By comparison, pSV3-neo was replicated in simian cells, but no recombined plasmids were detected by Southern analysis. No replication was detected after transfection of pSV3-neo into mouse cells. These results appear to reflect important differences between rabbit and simian cells in the efficiency of homologous recombination involving specific SV40 sequences and/or in the replication of the resulting recombinants. The role of the host cell in the evolution of SV40 sequences may influence the outcome of the infection with this virus.
携带SV40早期基因区域和新霉素抗性基因(neor)的质粒pSV3-neo,在转染到兔血管平滑肌细胞后,于小t抗原内含子的SV40重复序列与多聚腺苷酸化位点之间发生了同源重组。这种重组产生了两个分别为1.7 kb和3.0 kb的游离质粒,它们能在转染的兔细胞中复制,而载体质粒则不能。在由pSV3-neo转化的兔细胞中,这两个重组质粒持续复制了数月,其结构保持不变。在不同的pSV3-neo转化兔克隆中,发现每个细胞含有15至2510个拷贝的3.0 kb质粒以及370至1565个拷贝的1.7 kb质粒。将pSV3-neo转染到SV40转化的兔细胞3天后,也发现了类似的重组质粒。相比之下,pSV3-neo能在猴细胞中复制,但Southern分析未检测到重组质粒。将pSV3-neo转染到小鼠细胞后未检测到复制。这些结果似乎反映了兔细胞和猴细胞在涉及特定SV40序列的同源重组效率和/或所得重组体的复制方面存在重要差异。宿主细胞在SV40序列进化中的作用可能会影响该病毒感染的结果。