Role of cspA on the Preparation of Escherichia coli Competent Cells by Calcium Chloride Method.

Autor: Chen X; School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, China., Zhu N; School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, China.; School of Petrochemical Engineering, Lanzhou University of Technology, Lanzhou, China., Yang G; Gansu Zhongshang Food Quality Test and Detection Co. Ltd., Lanzhou, China.; Gansu Business Science and Technology Institute Co. Ltd., Lanzhou, China., Guo X; School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, China., Sun S; School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, China.; School of Petrochemical Engineering, Lanzhou University of Technology, Lanzhou, China., Leng F; School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, China., Wang Y; School of Life Science and Engineering, Lanzhou University of Technology, Lanzhou, China.
Jazyk: angličtina
Zdroj: Journal of basic microbiology [J Basic Microbiol] 2024 Aug; Vol. 64 (8), pp. e2400113. Date of Electronic Publication: 2024 Jun 25.
DOI: 10.1002/jobm.202400113
Abstrakt: One of the fundamental techniques in genetic engineering is the creation of Escherichia coli competent cells using the CaCl 2 method. However, little is known about the mechanism of E. coli competence formation. We have previously found that the cspA gene may play an indispensable role in the preparation of E. coli DH5α competent cells through multiomics analysis. In the present study, the cellular localization, physicochemical properties, and function of the protein expressed by the cspA gene were analyzed. To investigate the role of the cspA gene in E. coli transformation, cspA-deficient mutant was constructed by red homologous recombination. The growth, transformation efficiency, and cell morphology of the cspA-deficient strain and E. coli were compared. It was found that there were no noticeable differences in growth and morphology between E. coli and the cspA-deficient strain cultured at 37°C, but the mutant exhibited increased transformation efficiencies compared to E. coli DH5α for plasmids pUC19, pET-32a, and p1304, with enhancements of 2.23, 2.24, and 3.46 times, respectively. It was proved that cspA gene is an important negative regulatory gene in the CaCl 2 preparation of competent cells.
(© 2024 Wiley‐VCH GmbH.)
Databáze: MEDLINE
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