Confirmation of presence of recA306 mutation derived from polA deficient Esherichia coli isogenic cells with NGS sequencing in those long PCR product. Escherichia coli

Chromosome damage combined with defective recombinase activity has been widely considered to render cells inviable, owing to deficient double-strand break repair. However, temperature-sensitive recAts polA cells grow well upon induction of DNA damage and amelioration of ROS levels in those cells. Therefore, we investigated whether polA cells can tolerate a complete lack of recombinase function. We introduced a delta-recA allele in polA cells in series of isogenic E. coli cells which were differed in expression of redox chaperon. We confirmed presence of delta-recA mutation in those cells using Next-Generation Sequencing (NGS).