Isolation and characterization of novel cold-sensitive dnaA mutants of Escherichia coli

Lei Guo, Tsutomu Katayama, Yousuke Seyama, Kazuhisa Sekimizu, Takeyoshi Miki

研究成果: Contribution to journalArticle査読

11 被引用数 (Scopus)

抄録

We developed an efficient method for isolation of novel dnaA mutations based on PCR mutagenesis in the presence of manganese ion and shuffling of dnaA-carrying plasmids in a dnaA deletion host bacterium. Using this system, we obtained 30 cold-sensitive mutants from 4000 clones carrying plasmids with a mutagenized dnaA gene. All 27 cold-sensitive mutants analyzed were defective in DNA replication; none had a DnaAcos (over-initiation) phenotype. Nucleotide sequencing revealed that novel 15 alleles (mutations in 14 amino acid residues) are responsible for the cold-sensitive phenotype and are all located in the carboxy-terminal half of the DnaA protein. Copyright (C) 1999 Federation of European Microbiological Societies.

本文言語英語
ページ(範囲)357-366
ページ数10
ジャーナルFEMS microbiology letters
176
2
DOI
出版ステータス出版済み - 7 15 1999

All Science Journal Classification (ASJC) codes

  • 微生物学
  • 分子生物学
  • 遺伝学

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