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1型单纯疱疹病毒融合诱导MP株突变的精细定位

Fine mapping of mutations in the fusion-inducing MP strain of herpes simplex virus type 1.

作者信息

Pogue-Geile K L, Lee G T, Shapira S K, Spear P G

出版信息

Virology. 1984 Jul 15;136(1):100-9. doi: 10.1016/0042-6822(84)90251-4.

Abstract

Previous studies [W. T. Ruyechan, L. S. Morse, D. M. Knipe, and B. Roizman (1979) J. Virol. 29, 677-697] have shown that multiple mutations are responsible for the mutant phenotypes of herpes simplex virus type 1, strain MP, and have indicated that these mutations may be located on the genome between map coordinates 0.70 and 0.83. Strain MP produces large syncytial (Syn) plaques on many cell types and does not express glycoprotein C (gC-), whereas a sibling strain mP produces wild-type, small, nonsyncytial (Syn+) plaques and is gC+. Cloned DNA fragments from strains MP and mP (and strain F) were used in marker transfer and marker rescue experiments to map more precisely the mutations in MP. It was found that a 680-bp fragment from MP DNA (map coordinates 0.735 to 0.740) could transfer a Syn marker to mP and that, conversely, an overlapping fragment from mP DNA (map coordinates 0.728 to 0.744) could rescue the Syn mutation of MP. Recombinant viruses obtained in these experiments differed from the donor of the cloned DNA fragment in plaque size, however, indicating that mutation(s) at other regions of the MP genome cause enlarged plaques, in which the infected cells are less rounded than in wild-type plaques. A fragment of MP DNA from map coordinates 0.60 to 0.64 transferred a mutation causing the gC- phenotype to strain mP, and a fragment of F DNA from map coordinates 0.62 to 0.64 rescued the gC- mutation of MP. These results, coupled with data published by Frink et al. [(1983) J. Virol. 45, 643-467], indicate that the mutation responsible for the gC- phenotype of MP may be in the structural gene for gC.

摘要

先前的研究[W. T. 鲁耶尚、L. S. 莫尔斯、D. M. 克尼普和B. 罗伊兹曼(1979年)《病毒学杂志》第29卷,第677 - 697页]表明,多种突变导致了单纯疱疹病毒1型MP株的突变表型,并指出这些突变可能位于基因组图谱坐标0.70至0.83之间。MP株在许多细胞类型上产生大型合胞体(Syn)噬斑,且不表达糖蛋白C(gC-),而其同源株mP产生野生型、小型、非合胞体(Syn+)噬斑,且是gC+。来自MP株和mP株(以及F株)的克隆DNA片段用于标记转移和标记拯救实验,以更精确地定位MP株中的突变。结果发现,来自MP DNA的一个680碱基对片段(图谱坐标0.735至0.740)可将Syn标记转移至mP株,相反,来自mP DNA的一个重叠片段(图谱坐标0.728至0.744)可拯救MP株的Syn突变。然而,在这些实验中获得的重组病毒在噬斑大小上与克隆DNA片段的供体不同,这表明MP基因组其他区域的突变导致噬斑增大,其中感染细胞比野生型噬斑中的细胞更不呈圆形。来自图谱坐标0.60至0.64的MP DNA片段将导致gC-表型的突变转移至mP株,来自图谱坐标0.62至0.64的F DNA片段拯救了MP株的gC-突变。这些结果,再加上弗林克等人发表的数据[(1983年)《病毒学杂志》第45卷,第643 - 467页],表明导致MP株gC-表型的突变可能位于gC的结构基因中。

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