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人腺病毒2型温度敏感突变体的分离与表型特征分析

Isolation and phenotypic characterization of human adenovirus type 2 temperature-sensitive mutants.

作者信息

Martin G R, Warocquier R, Cousin C, D'Halluin J C, Boulanger P A

出版信息

J Gen Virol. 1978 Nov;41(2):303-14. doi: 10.1099/0022-1317-41-2-303.

Abstract

Thirty-nine temperature-sensitive (ts) mutants that fail to grow at 39.5 degrees C but develop normally at 33 degrees C have been isolated from a nitrous-acid-treated stock of a wild-type strain of type 2 human adenovirus. The frequency of ts mutants among the surviving viruses was about 10%. Complementation tests in doubly infected cell cultures at restrictive temperature permitted the assignment of 19 of these mutants to 11 complementation groups. They are characterized phenotypically according to their soluble capsid antigen production quantified by two-dimensional immunoelectrophoresis, virus DNA synthesis, as measured by alkaline sucrose gradient sedimentation of 34S DNA, and virion morphogenesis, as analysed by electron microscopy os cell sections. Two complementation groups were defective for DNA synthesis, four for soluble hexon production and two groups for total penton (penton base + fibre), while one group revealed no fibre production. Two complementation groups presented a normal antigen pattern, but the particles exhibited altered morphology as observed in cell sections.

摘要

从经亚硝酸处理的2型人类腺病毒野生型菌株的储备中分离出了39个温度敏感(ts)突变体,这些突变体在39.5℃时无法生长,但在33℃时能正常发育。存活病毒中ts突变体的频率约为10%。在限制温度下对双重感染的细胞培养物进行互补试验,可将其中19个突变体归为11个互补组。根据通过二维免疫电泳定量的可溶性衣壳抗原产生、通过34S DNA的碱性蔗糖梯度沉降测量的病毒DNA合成以及通过细胞切片电子显微镜分析的病毒粒子形态发生,对它们进行表型特征描述。两个互补组在DNA合成方面有缺陷,四个在可溶性六邻体产生方面有缺陷,两个组在总五邻体(五邻体基部+纤维)方面有缺陷,而一个组没有纤维产生。两个互补组呈现正常的抗原模式,但在细胞切片中观察到粒子形态发生了改变。

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