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31型腺病毒左端DNA片段转化的大鼠细胞系的建立与鉴定

Establishment and characterization of rat cell lines transformed by the left-end DNA fragments of adenovirus type 31.

作者信息

Yamashita T, Fujinaga K

出版信息

Gan. 1983 Feb;74(1):77-85.

PMID:6840439
Abstract

A rat cell line, 3Y1, was transfected with the left-end DNA fragments of adenovirus (Ad) 31 DNA and transformed cell lines were established. A cell line 31BY4-1, which was induced by Ad31 BamHI-B (left-most 17.4% of the genome), showed typical transformation phenotypes, while cell lines 31GY1-2, 31GY2-5, and 31GY3-4, which were induced by Ad31 HindIII-G (left-most 6.7% of the genome), showed intermediate phenotypes between transformed and untransformed cells, with the following properties: (1) percent plating efficiencies in Eagle's minimum essential medium with 2% fetal calf serum and in soft-agar culture were extremely low, like those of 3Y1 cells; (2) they form tumors in newborn rats only after long latent periods. Southern blot hybridization revealed that, in all the transformed cell lines, viral DNA sequences were integrated at multiple loci in large-molecular-weight cell DNAs. Northern blot hybridization showed that viral mRNAs of early region 1A (E1A) and early region 1B (E1B) were both transcribed in 31BY4-1 cells as well as in KB cells early after infection with Ad31. On the other hand, E1B viral mRNA was present in a much lesser amount in 31GY cell lines, or was undetectable.

摘要

用腺病毒(Ad)31 DNA的左端DNA片段转染大鼠细胞系3Y1,并建立了转化细胞系。由Ad31 BamHI - B(基因组最左端的17.4%)诱导产生的细胞系31BY4 - 1表现出典型的转化表型,而由Ad31 HindIII - G(基因组最左端的6.7%)诱导产生的细胞系31GY1 - 2、31GY2 - 5和31GY3 - 4表现出介于转化细胞和未转化细胞之间的中间表型,具有以下特性:(1)在含2%胎牛血清的伊格尔最低必需培养基中以及软琼脂培养中的铺板效率百分比极低,与3Y1细胞相似;(2)它们仅在长时间潜伏期后才在新生大鼠中形成肿瘤。Southern印迹杂交显示,在所有转化细胞系中,病毒DNA序列整合到高分子量细胞DNA的多个位点。Northern印迹杂交表明,在感染Ad31后早期,早期区域1A(E1A)和早期区域1B(E1B)的病毒mRNA在31BY4 - 1细胞以及KB细胞中均有转录。另一方面,E1B病毒mRNA在31GY细胞系中的含量要少得多,或者无法检测到。

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