A study on creating an attenuated Aeromonas hydrophila mutant strain by knocking out aroA gene

Truong Ngoc Thuy Lien, Vu Thi Thanh Huong, Tran Thanh Tieng, Nguyen Quoc Binh

Abstract


In order to develop live attenuated vaccine against Aeromonas hydrophila infection in fish, Aeromonas hydrophila mutant strain was created by knocking out the aroA gene, encoding 5-enolpyruvylshikimate 3-phosphate synthase. A 683 bp middle fragment of this gene was replaced by kanamycin resistance gene KmR to make the cassette aroA::KmR. The E. coli Sm10 λpir strain  carrying suicide pGP704 plasmid with aroA::KmR was used as donor in conjugation with A. hydrophila wild type to create the mutant. The toxicity of the mutant strain was determined by intra-peritoneal injection into Tra catfish fingerlings. The results showed that the LD50 of the mutant strain was >107 CFU/ml, whereas that of the wild type strain was <103 CFU/ml

Keywords


Aeromonas hydrophila, attenuated, knockout gene, mutant, vaccine.

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DOI: https://doi.org/10.15625/0866-7160/v36n1se.4335 Display counter: Abstract : 49 views. PDF : 85 views.

 

                 

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