Expression of herpes simplex virus glycoprotein C from a DNA fragment inserted into the thymidine kinase gene of this virus.
Expression of herpes simplex virus glycoprotein C from a DNA fragment inserted into the thymidine kinase gene of this virus.
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单纯疱疹病毒糖蛋白 C 从插入该病毒胸苷激酶基因的 DNA 片段中表达。
DOI：
10.1073/pnas.79.21.6612
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发表时间：
1982
影响因子：
11.1
通讯作者：
Spear,PG
中科院分区：
文献类型：
--
作者：
Lee,GT;Pogue-Geile,KL;Pereira,L;Spear,PG
以前的报道已经描述了1型单纯疱疹病毒的突变体，其不能产生或积累主要糖蛋白之一，糖蛋白C（gC）。这种缺陷在细胞培养中是不致命的，与一些突变体的合胞体斑块形态有关，并且可能是由映射到基因组上与gC结构基因不相邻的区域的突变引起的。为了研究在特定遗传背景下gC表达所需的条件和后果，我们将gC基因的野生型等位基因插入到gC融合诱导病毒突变株MP的胸苷激酶基因（tk）中。这是通过鉴定与gC mRNA同源的克隆病毒DNA片段，将适当的片段插入克隆在pBR 322中的病毒tk中，然后用重组质粒和来自MP株的DNA共转染细胞，以选择插入TK-突变体来实现的。发现所有含有插入适当序列（以任一方向）到tk中的TK-突变体表达gC，同时保持菌株MP的合胞体噬斑形态。从一个TK-突变体中消除插入伴随着产生gC的能力的丧失。我们的结果允许编码gC的DNA序列更精确地定位到Sal I片段R的亚片段（图坐标0.620-0.640），并且还表明gC基因的启动子序列可能位于该片段中。此外，我们可以得出结论，先前描述的调节突变的菌株MP不阻止gC的表达从插入到其基因tk的DNA和MP的合胞体表型不能仅仅是由于缺乏gC。
Previous reports have described mutants of herpes simplex virus type 1 that fail to produce or accumulate one of the major glycoproteins, glycoprotein C (gC). This defect is not lethal in cell culture, has been associated with the syncytial plaque morphology of some mutants, and may result from mutations that map to a region on the genome noncontiguous with the structural gene for gC. To investigate the conditions required for, and consequences of, gC expression in a specific genetic background, we have inserted a wild-type allele of the gC gene into the thymidine kinase gene (tk) of a gC- fusion-inducing viral mutant, strain MP. This was accomplished by identifying cloned viral DNA fragments homologous to gC mRNA, inserting the appropriate fragments into the viral tk cloned in pBR322, and then cotransfecting cells with the recombinant plasmids and DNA from strain MP, for selection of insertional TK- mutants. All TK- mutants containing insertions of appropriate sequences (in either orientation) into tk were found to express gC while maintaining the syncytial plaque morphology of strain MP. Elimination of the insertion from one of the TK- mutants was accompanied by loss of ability to produce gC. Our results permit more precise mapping of the DNA sequence encoding gC, to a subfragment of Sal I fragment R (map coordinates 0.620-0.640) and indicate also that promoter sequences for the gC gene may be located in this fragment. Moreover, we can conclude that the previously described regulatory mutation of strain MP does not prevent expression of gC from the DNA inserted into its gene tk and that the syncytial phenotype of MP cannot be due solely to absence of gC.