Expression and characterization of the rabies virus glycoprotein in mammalian system Krithiga N1, Bokade Prajakta P2, Balamurugan V2, Gupta P3, Babu Aravindh RP4, Rathnamma D1, Sharada R1, Dilip L1, Krupa KN1, Hegde Nagendra R4, Isloor S1,* 1Department of Microbiology, Veterinary College, KAVAFSU, Hebbal, Bengaluru, Karnataka, India 2ICAR-NIVEDI, Yelahanka, Bengaluru, Karnataka, India 3National Institute of Animal Biotechnology, Hyderabad, Telangana, India 4TRPVB, TANUVAS, Chennai, Tamil Nadu, India *Corresponding author E-mail id: kisloor@gmail.com
Onilne Published on 8 March, 2024. Abstract The utilization of heterologous gene expression technology was one of several methods employed to generate novel rabies vaccines with enhanced properties compared to the currently existing vaccines. Hence, the current study aimed at cloning and expressing the rabies virus glycoprotein (RABG) in the mammalian system. The study targeted the cloning of the ectodomain of RABG in the pTriEX-2 mammalian expression vector, followed by transient transfection using the synthetic polymer polyethylenimine (PEI) in Human Embryonic Kidney (HEK 293) cells. The study confirmed the successful construction of RABG ectodomain by colony PCR, restriction digestion, and gene sequencing. Subsequent expression profiling of RABG was performed through Reverse Transcriptase PCR (RT-PCR). The RT-PCR confirmed the expression of RABG in HEK293 by the existence of mRNA transcripts of RABG from the recombinant clones. The present RABG construct might be useful for the development of newer diagnostic techniques or, with further research, it can lay the groundwork for the development of a more effective recombinant vaccine for canine rabies. Top Keywords Rabies, Glycoprotein, Ectodomain, pTriEX-2, HEK, Expression. Top |