Related Experiment Video
Updated: May 22, 2026

Laboratory Techniques Used to Maintain and Differentiate Biotypes of Vibrio cholerae Clinical and Environmental Isolates
Published on: May 30, 2017
[Isolation and function of genes regulating aphB expression in Vibrio cholerae]
Haili Chen1, Zhaoqin Zhu, Zengtao Zhong
1Pathogen Diagnosis and Biosafety Department, Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China. hai_li_c@sina.com
Objective:
We identified genes that regulate the expression of aphB, the gene encoding a key virulence regulator in Vibrio cholerae O1 E1 Tor C6706(-).
Methods:
We constructed a transposon library in V. cholerae C6706 strain containing a P(aphB)-luxCDABE and P(aphB)-lacZ transcriptional reporter plasmids. Using a chemiluminescence imager system, we rapidly detected aphB promoter expression level at a large scale. We then sequenced the transposon insertion sites by arbitrary PCR and sequencing analysis.
Results:
We obtained two candidate mutants T1 and T2 which displayed reduced aphB expression from approximately 40,000 transposon insertion mutants. Sequencing analysis shows that Tn inserted in vc1585 reading frame in the T1 mutant and Tn inserted in the end of coding sequence of vc1602 in the T2 mutant.
Conclusion:
By using a genetic screen, we identified two potential genes that may involve in regulation of the expression of the key virulence regulator AphB. This study sheds light on our further investigation to fully understand V. cholerae virulence gene regulatory cascades.
Related Concept Videos
Regulation of Bacterial Virulence
Coordination of Gene Expression Processes in Bacteria
Cholera
Other Stress Responses in Bacteria
Gram-negative Bacterial Protein Secretion Systems
Stringent Response in E. coli

