Vet World   Vol.10   September-2017  Article-14

Cloning and sequence analysis of hyaluronoglucosaminidase (nagH) gene of Clostridium chauvoei

Background and Aim: Blackleg disease is caused by Clostridium chauvoei in ruminants. Although virulence factors such as C. chauvoei toxin A, sialidase, and flagellin are well characterized, hyaluronidases of C. chauvoei are not characterized. The present study was aimed at cloning and sequence analysis of hyaluronoglucosaminidase (nagH) gene of C. chauvoei.

Materials and Methods: C. chauvoei strain ATCC 10092 was grown in ATCC 2107 media and confirmed by polymerase chain reaction (PCR) using the primers specific for 16-23S rDNA spacer region. nagH gene of C. chauvoei was amplified and cloned into pRham-SUMO vector and transformed into Escherichia cloni 10G cells. The construct was then transformed into E. cloni cells. Colony PCR was carried out to screen the colonies followed by sequencing of nagH gene in the construct.

Results: PCR amplification yielded nagH gene of 1143 bp product, which was cloned in prokaryotic expression system. Colony PCR, as well as sequencing of nagH gene, confirmed the presence of insert. Sequence was then subjected to BLAST analysis of NCBI, which confirmed that the sequence was indeed of nagH gene of C. chauvoei. Phylogenetic analysis of the sequence showed that it is closely related to Clostridium perfringens and Clostridium paraputrificum.

Conclusion: The gene for virulence factor nagH was cloned into a prokaryotic expression vector and confirmed by sequencing. Keywords: black quarter, Clostridium chauvoei, hyaluronoglucosaminidase.

Keywords: black quarter, Clostridium chauvoei, hyaluronoglucosaminidase.

How to cite this article: Dangi SK, Yadav PK, Tiwari A, Nagaleekar VK (2017) Cloning and sequence analysis of hyaluronoglucosaminidase (nagH) gene of Clostridium chauvoei, Veterinary World, 10(9): 1104-1107.

Received: 28-07-2017  Accepted: 11-08-2017     Published online: 21-09-2017

Corresponding author: Viswas Konasagara Nagaleekar   E-mail: vkn111@gmail.com

DOI: 10.14202/vetworld.2017.1104-1107

Copyright: Dangi, et al. This article is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http:// creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.