Efficient construction of Haemophilus parasuis mutants based on natural transformation
Studies on virulence factors and pathogenecity of Haemophilus parasuis have long been hindered by a lack of a consistent system for genetic manipulation. In this study, competence was induced by transferring H. parasuis from rich medium to starvation medium media-IV (M-IV)
and iscR gene deficient mutants of H. parasuis were generated efficiently. Transformation frequency varied from 4.1 × 10−5 to 1.1 × 10−8 when using circular plasmid, and increased to about 2- to 31-fold when transformed using linearized
plasmid. Allele replacement occurred efficiently in 6 strains, which are transformable using both circular and linearized pTRU, but not in another 2 strains which could only be transformed using linearized plasmid. The iscR mutants were stable for at least 20 passages in vitro. Haemophilus
parasuis strains vary extensively in natural transformation efficiency and the method established here allows for transformation of a larger spectrum of strains with an easily accessed plasmid. This provides important tools for genetic manipulation of H. parasuis.
Document Type: Research Article
Publication date: 01 October 2016
- The Canadian Journal of Veterinary Research (CJVR), published by Canadian Veterinary Medical Association, is Canada's only veterinary research publication. This quarterly peer-reviewed journal has earned a wide international readership through the publishing of high quality scientific papers in the field of veterinary medicine. CJVR publishes the results of original research in veterinary and comparative medicine.
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