Development of Live Vaccines from Yersinia Ruckeri by Aroa and Aroc Gene Mutations
Razvijanje vakcine protiv yersinia ruckeri pomoću aroa i aroc genske mutacije
Author
Altinok, Ilhan
Kahraman, Umit
Capkin, Erol
Boran, Halis
Ozturk, Rafet
Keywords
Aromatic amino acid biosynthesis network (aro)aroA
aroC
overlap PCR
live attenuated vaccine
sinteza aromatičnih amino kiselina
živa atenuisana vakcina
Metadata
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Yersiniosis caused by Yersinia ruckeri is the main causes of high mortalities and severe economic losses in freshwater and marine aquaculture. To prevent this disease, a live attenuated vaccine was developed. For this purpose, Y. ruckeri aroA and aroC genes were amplified and DNA fragments mutated by overlap extension PCR was cloned into a suicide plasmid. This plasmid was transferred to Y. ruckeri for replacing wild type genes with mutated aroA and aroC genes via homologous recombination. Virulence of mutant Y. ruckeri was determined by infecting juvenile rainbow trout with mutant bacteria and calculating fish mortalities and immune responses in fish. Similarly, efficacy of mutant Y. ruckeri was determined by vaccinating fish with mutant bacteria and then challenging the same fish with wild type Y. ruckeri RB0708 6 months after vaccination. Vaccination of rainbow trout with the aroA and aroC mutant as a live vaccine conferred significant protection against the wild-type Y. ruckeri. Jersinioza koju izaziva Yersinia ruckeri je osnovni uzrok visokog mortaliteta i velikih ekonomskih gubitaka u slatkovodnoj i marinskoj akvakulturi. Da bi se bolest sprečila razvijena je živa atenuisana vakcina. U tu svrhu su geni Y. ruckeri aroA i aroC umnoženi i DNK fragmenti mutirani korišćenjem PCR metode kojom su klonirani i ubačeni u plazmid. Plazmid je prenet u Y. ruckeri da bi zamenio divlji tip gena mutiranim aroA i aroC genima, putem homologe rekombinacije. Virulenca mutiranog Y. ruckeri je određivana inficiranjem mlađi kalifornijske pastrmke mutiranim bakterijama i izračunavanjem mortaliteta i imunog odgovora riba. Efikasnost mutiranog Y. ruckeri je određivana vakcinacijom riba mutiranim bakterijama i izazivanjem obolenja kod iste ribe divljim tipom Y. ruckeri RB0708 6 meseci po vakcinaciji. Vakcinacija kalifornijske pastrmke sa aroA i aroC mutantima kao žive vakcine doprineo je značajnoj zaštiti protiv divljeg soja Y. ruckeri.