Bioinformatics analysis of gene bhsA and its role in Ca2+‐treated Escherichia coli

Abstract One of the commonly employed methods in molecular biology is to utilize calcium chloride to treat Escherichia coli for the preparation of competent cells to facilitate foreign gene expression. However, the molecular mechanisms underlying Ca 2+ mediation of competent cell formation and identification of the key genes involved in the process remain unclear. In previous studies, the combined analysis of transcriptomics and proteomics revealed bhsA as one of the crucial genes. The gene ontology functional annotation of bhsA identified it as a member of the YhcN family encoding an outer membrane protein that confers resistance to various stresses. The IPR0108542 domain found within the protein plays a significant role in stress response and biofilm formation in E. coli . Analysis of the STRING database showed that the proteins interacting with bhsA are primarily involved in biofilm formation and stress resistance. Using the RED homologous recombination method, a bhsA deletion strain was constructed to verify its role in E. coli genetic transformation. Although the mutant strain showed no significant differences in morphology or growth trend when compared to the wild‐type strain, its transformation efficiency decreased by 1.14‐ and 1.64‐fold with plasmids pUC19 and pET‐32a. Furthermore, the 1‐ N ‐phenylnaphthylamine assay indicated a 1.71‐fold reduction in cell membrane permeability in the mutant strain.