[Construction of recombinant E. coli-L. interrogans shuttle plasmid pGKINVA and screening recombinant leptospira strains].
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.


CONCLUSIONS
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.
Zhu
Qing-ping
QP
Unit of Infection and Immunity, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China
Bao
Lang
L
Zhang
Hui-dong
HD
Zhao
Ming-cai
MC
D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification; D004274:DNA, Recombinant / Q000235:genetics*; D004586:Electrophoresis; D004926:Escherichia coli / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D007921:Leptospira interrogans / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction
D001426:Bacterial Proteins; D004274:DNA, Recombinant; C075518:invA protein, Bacteria