Cloning and expression of partial CDs of leptospiral beta propeller repeat protein in prokaryotic system Manoharan Balumahendiran1,*, Devalam Rani Prameela2, Bollini Sreedevi3, Rao Vaikunta V.4, Babu Jagadeesh A.5 1Faculty of Veterinary Sciences, Tamil Nadu Agricultural University, Coimbatore, Tamil Nadu, India 2State Level Diagnostic Laboratory, Sri Venkateswara Veterinary Univeristy, Tirupati, Andhra Pradesh, India 3College of Veterinary Science, Gannavaram, Sri Venkateswara Veterinary Univeristy, Tirupati, Andhra Pradesh, India 4College of Veterinary Science, Proddatur, Sri Venkateswara Veterinary Univeristy, Tirupati, Andhra Pradesh, India 5College of Veterinary Science, Sri Venkateswara Veterinary Univeristy, Tirupati, Andhra Pradesh, India *Corresponding author: M Balumahendiran; E-mail: balumb@gmail.com
Online Published on 30 December, 2022. Abstract Leptospirosis is the wide spread neglected zoonotic disease among domestic and pet animals and poses major threat to human public health. Although it is treatable with antibiotics, vaccination is the prime strategy to control the disease. However, the current vaccines are ineffective, lack of cross reactivity and require booster doses. Therefore, it is inevitable to identify conserved protein candidates that can provide immunity against the majority of serovars. In this context, the present study was aimed to clone and express the lipoprotein Beta Propeller repeat (BPR) of L. canicola. In this study, 435 base pair of partial coding sequences between 195 to 340 aa region of beta propeller gene was amplified by PCR. The fragment of 435 bp BPR gene was inserted into the pET 32(a) vector and expressed in E. coli Bl21(DE3) cells. SDS-PAGE revealed an expected size of 36 kDa and the immunoblot with anti sera raised against the whole cell lysate of L. canicola, confirmed the specificity of the protein expressed in E. coli system. NCBI BLAST analysis showed that 435 nucleotides are flanking the coding region of 422164 to 422598 positions of L. interrogans sps chromosome number 1 and having 95 percent identities with the published sequences. Phylogenetic analysis revealed that the BPR gene of this study occupied the same clade in the phylogenetic tree as other L. canicola serovars. From the findings of this study, it may be concluded that the Beta Propeller Repeat protein gene was conserved in the genus Leptospira and which could be a potential vaccine candidate for subunit vaccines. Highlights • Beta propeller repeat protein of L. canicola was amplified, sequenced partially and expressed in prokaryotic system. • BLAST analysis showed that 435 nucleotides are flanking the coding region of 422164 to 422598 positions of L. interrogans spp. • Immuno blot and in-silico analysis confirmed its specificity and conservancy among other serovars of Leptospira sps Top Keywords Beta Propeller Repeat gene, Canine Leptospirosis, PCR, Cloning, Expression. Top |