Construction and Characterization of purD Gene Deleted Brucella abortus
Construction and Characterization of purD Gene Deleted Brucella abortus
Muhammad Ilyas Riaz1, Masood Rabbani1*, Sohail Raza1, Ali Raza Awan2, Aleena Kokab1 and Iqra Nazir3
ABSTRACT
Bovine brucellosis is an important zoonotic and contagious disease targeting humans, domestic and wild animals all over the globe resulting in significant economic losses therefore, disease mitigation and control measures are of pronounced significance. The two available vaccines of Brucella abortus in Pakistan, RB51, and S19 have many limitations in the light of residual virulence, induction of abortion, diagnostic interference, and human pathogenic potential. In this study, we have developed a Brucella abortus pUC19-K-UP-DN plasmid cassette for the construction of ΔpurD deleted mutant by deleting the purD gene which is responsible for de novo purine synthesis. For this, the pUC19-K-UP-DN plasmid cassette was initially constructed using pUC19, kanamycin, and purD upstream and downstream fragments. Then the cassette was electroporated into electro competent Brucella abortus RB51. The ΔpurD mutant was confirmed using PCR analysis. Afterward, the growth kinetics of the mutant was compared with parent RB51. The mutant bacterial growth significantly reduces (P< 0.05) as compared to the parent non-mutant confirming its auxotroph inability towards purine pathway biosynthesis which serves as a crucial factor for disease induction and survival inside the host system revealing enhanced attenuation in the deleted mutant. However, after supplementation of purine bases, the ΔpurD mutant bacteria regain growth in enriched media. The results confirmed that the highly attenuated Brucella abortus ΔpurD mutant can be used successfully as a potential vaccine candidate for the control of bovine brucellosis.
To share on other social networks, click on any share button. What are these?
