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小鼠巨细胞病毒基因组HindIII J片段中的基因对于在培养细胞中的生长并非必需:利用lacZ/gpt盒进行插入诱变

Genes in the HindIII J fragment of the murine cytomegalovirus genome are dispensable for growth in cultured cells: insertion mutagenesis with a lacZ/gpt cassette.

作者信息

Vieira J, Farrell H E, Rawlinson W D, Mocarski E S

机构信息

Department of Microbiology and Immunology, Stanford University School of Medicine, California 94305-5402.

出版信息

J Virol. 1994 Aug;68(8):4837-46. doi: 10.1128/JVI.68.8.4837-4846.1994.

Abstract

The organization and function of the genes encoded within the HindIII J region of the murine cytomegalovirus genome were analyzed by transcript mapping and cDNA isolation, nucleotide sequence analysis and identification of open reading frames (ORFs), and construction of recombinant viruses carrying insertions disrupting five of the seven ORFs. This region was found to encode five beta transcripts and one gamma transcript in addition to two beta transcripts previously mapped to the sgg1 locus. Seven open reading frames were identified, and one was recognized as a homolog of a human cytomegalovirus US22 gene family. The five largest ORFs contained within the HindIII J fragment (sgg1, HJ4, HJ5, HJ6, and HJ7) were each disrupted by the insertion of a lacZ/gpt genetic marker cassette. The growth kinetics of all recombinant viruses were investigated and found to be the same as wild-type parental virus, indicating that these five ORFs were dispensable for growth in cell culture.

摘要

通过转录图谱分析和cDNA分离、核苷酸序列分析以及开放阅读框(ORF)的鉴定,以及构建携带插入片段破坏七个ORF中的五个的重组病毒,对鼠巨细胞病毒基因组HindIII J区域内编码的基因的组织和功能进行了分析。除了先前定位到sgg1位点的两个β转录本外,该区域还被发现编码五个β转录本和一个γ转录本。鉴定出七个开放阅读框,其中一个被认为是人类巨细胞病毒US22基因家族的同源物。HindIII J片段(sgg1、HJ4、HJ5、HJ6和HJ7)中包含的五个最大的ORF各自通过插入lacZ/gpt遗传标记盒而被破坏。研究了所有重组病毒的生长动力学,发现其与野生型亲本病毒相同,表明这五个ORF对于细胞培养中的生长是可有可无的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/38aa/236423/f8d1332d2e73/jvirol00017-0145-a.jpg

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