Cloning and sequence analysis of hyaluronoglucosaminidase gene of .

AIM

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

MATERIALS AND METHODS

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. gene of was amplified and cloned into pRham-SUMO vector and transformed into 10G cells. The construct was then transformed into cells. Colony PCR was carried out to screen the colonies followed by sequencing of gene in the construct.

RESULTS

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

CONCLUSIONS

The gene for virulence factor was cloned into a prokaryotic expression vector and confirmed by sequencing.