Shuttle vector system for the analysis of mutational events in mammalian chromosomal DNA

Hironobu Ikehata, Hiroshi Kimura, Takesi Kato

研究成果: Article査読

11 被引用数 (Scopus)

抄録

cDNA of the human hprt gene was introduced into the BamHI cloning site of the retroviral shuttle vector pZipNeoSV(X)1. The mouse cell line 2TGOR, a hypoxanthine phosphoribosyltransferase-deficient derivative of Balb/c 3T3, was tranformed with the vector and some stably transformed HATrNEOr clones were established. One of the clones, VH-12, contained a single copy of the vector integrated stably into a chromosome in a proviral form. From this clone, we were able to recover efficiently the vector sequence preserving its intact strucure by use of COS cell fusion. The relatively small size of the hprt cDNA (657 base pairs for the coding region) allowed quick determination of the entire DNA sequence. It was also notable that use of 6TG NEO double selection for mutant isolation could eliminate the 6TGr derivatives of VH-12 cells which arose from loss of the total vector sequence or from some epigenetic event, because such alterations would lead to inactivation of the neo gene as well as the hprt cDNA. The properties of our shuttle vector system were particularly useful for analysis of the molecular mechanisms of mutational events in chromosomal DNA of mammalian cells.

本文言語English
ページ(範囲)237-247
ページ数11
ジャーナルMutation Research - Fundamental and Molecular Mechanisms of Mutagenesis
210
2
DOI
出版ステータスPublished - 1989 2
外部発表はい

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Health, Toxicology and Mutagenesis

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