著者

Shingo Suzuki Akihiro Kaidow Takuma Meya Anri Masuya Takashi Shiina

出版者

公益財団法人 応用微生物学・分子細胞生物学研究奨励会

雑誌

The Journal of General and Applied Microbiology (ISSN:00221260)

巻号頁・発行日

vol.63, no.1, pp.22-27, 2017 (Released:2017-03-17)

参考文献数

38

被引用文献数

2

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Many significant gene mutations in E. coli have contributed to the development of genetics. Among these, a commonly used recA mutation, Δ(srl–recA)306 has been sequenced by a next-generation sequencer combined with a long PCR. An original report described that Δ(srl–recA)306 cells were deleted from srlR to recA genes in their genome. The next-generation sequencer enables more accurate details to be determined. We ask whether both surrounding genes from hypF to norV for srlR and alaS for recA is there first. The long PCR was carried out with primers, norR and alaS, and amplified DNA fragments differed in length from wild to Δ(srl–recA)306 cells, suggesting that an entire Δ(srl–recA)306 mutation was included. Sequences of those DNA fragments indicated that 9147 bp, from srlR to recA including 10 genes, were replaced by a Tn10 DNA sequence. Junction points at both srlR-Tn10 and Tn10-recA were determined precisely. The results indicate that the first 97% of recA gene sequences were lost with a downstream recX gene remaining intact. The phenotypic difference between Δ(srl–recA)306 and ΔrecA::Km is discussed.