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在碘脱氧尿苷诱导的C3H/MuLV细胞体外转化的上皮性水貂肺细胞中,水貂细胞灶形成型鼠白血病病毒的基因组结构

Genome structure of mink cell focus-forming murine leukemia virus in epithelial mink lung cells transformed vitro by iododeoxyuridine-induced C3H/MuLV cells.

作者信息

Rapp U R, Birkenmeier E, Bonner T I, Gonda M A, Gunnell M

出版信息

J Virol. 1983 Feb;45(2):740-54. doi: 10.1128/JVI.45.2.740-754.1983.

Abstract

We characterized mink cell focus-forming murine leukemia viruses that were isolated from C3H/MCA-5 cells after induction with 5-iododeoxyuridine in culture. Mink lung epithelial cells malignantly transformed in vitro by induced virus were the source of four molecular clones of mink cell focus-forming virus. CI-1, CI-2, CI-3, and CI-4. Three clones, CI-1, CI-2, and CI-3, had full-length mink cell focus-forming viral genomes, one of which (CI-3) was infectious. In addition, we obtained a defective viral genome (CI-4) which had a deletion in the envelope gene. A comparison between the envelope genes of CI-4 and those of spleen focus-forming virus by heteroduplex mapping showed close homology in the substitution region and defined the deletion as being identical to the p15E deletion of spleen focus-forming virus. The recombinant mink cell focus-forming genomes are not endogenous in C3H/MCA-5 cells and therefore must have been formed in culture after induction by 5-iododeoxyuridine. CI-3, the infectious clone of mink cell focus-forming murine leukemia virus, was dualtropic, and mink cells infected with CI-3 were altered in their response to epidermal growth factor. In the presence of epidermal growth factor at 10 ng/ml, uninfected mink cells retained their epithelial morphology in monolayer culture and did not form colonies in soft agar. In contrast, CI-3 virus-infected mink cells grew with fibroblastic morphology in monolayer culture and showed an increased growth rate in soft agar in the presence of epidermal growth factor.

摘要

我们对在培养过程中用5-碘脱氧尿苷诱导后从C3H/MCA-5细胞中分离出的水貂细胞灶形成性鼠白血病病毒进行了特性分析。由诱导病毒在体外恶性转化的水貂肺上皮细胞是水貂细胞灶形成病毒的四个分子克隆CI-1、CI-2、CI-3和CI-4的来源。三个克隆CI-1、CI-2和CI-3具有全长水貂细胞灶形成病毒基因组,其中一个(CI-3)具有感染性。此外,我们获得了一个有缺陷的病毒基因组(CI-4),其包膜基因存在缺失。通过异源双链图谱比较CI-4的包膜基因与脾灶形成病毒的包膜基因,发现在替换区域有密切的同源性,并确定该缺失与脾灶形成病毒的p15E缺失相同。重组的水貂细胞灶形成基因组在C3H/MCA-5细胞中不是内源性的,因此一定是在5-碘脱氧尿苷诱导后在培养过程中形成的。水貂细胞灶形成性鼠白血病病毒的感染性克隆CI-3具有双嗜性,感染CI-3的水貂细胞对表皮生长因子的反应发生了改变。在10 ng/ml表皮生长因子存在的情况下,未感染的水貂细胞在单层培养中保持其上皮形态,在软琼脂中不形成集落。相比之下,感染CI-3病毒的水貂细胞在单层培养中以成纤维细胞形态生长,并且在表皮生长因子存在的情况下在软琼脂中显示出增加的生长速率。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3410/256469/2c230c50f71c/jvirol00149-0266-a.jpg

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