Related Experiment Video
Updated: Jul 17, 2026
Application of CRISPR Interference (CRISPRi) for Gene Silencing in Pathogenic Species of Leptospira
Published on: August 14, 2021
[Construction of recombinant E. coli-L. interrogans shuttle plasmid pGKINVA and screening recombinant leptospira
Qing-ping Zhu1, Lang Bao, Hui-dong Zhang
1Unit of Infection and Immunity, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China
Objective:
To further investigate pathogenic function of invA in Leptospira, recombinant L. biflexa-Escherichia coli shuttle vector pGKINVA was constructed, and electropored into L. biflexa strain Patoc I .
Methods:
The invA gene was cloned into shuttle vector pGKble24 to construct a recombinant L. biflexa-Escherichia coli shuttle vector pGKINVA. The recombinant vectors were electropored into L. biflexa serovar Patoc strain Patoc I .
Results:
A 558 bp invA was amplified from L. interrogans serovars Lai strain 017 and ligated with Aat II and Xma I digested pGKble24. After screening by kanamycin and blue/white color, the recombinant pGKINVA was obtained and sequenced. Subsequently, the recombinant pGKINVA was electropored into L. biflexa strain Patoc I. The selected mutants were identified by PCR.
Conclusion:
Recombinant L. biflexa-Escherichia coli shuttle vector with invA insert was constructed and two strains of pGKINVA transformed Leptospira were obtained. These pGKINVA transformed Leptospira strains would provid an experimental model to investigate potential pathogenic functions of invA in vivo.
More Related Videos
08:09Peptide Scanning-assisted Identification of a Monoclonal Antibody-recognized Linear B-cell Epitope
Published on: March 24, 2017
11:50High-throughput Parallel Sequencing to Measure Fitness of Leptospira interrogans Transposon Insertion Mutants During Golden Syrian Hamster Infection
Published on: December 18, 2017