Molecular characterization and in-silico analysis of complete coding sequence of bubaline mLYS

NIHAR RANJAN SAHOO; PUSHPENDRA KUMAR; T K BHATTACHARYA; A P SINGH; B BHUSHAN; A SHARMA; A K TIWARI

doi:10.56093/ijans.v85i12.54366

Authors

NIHAR RANJAN SAHOO Scientist, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 243 122 India
PUSHPENDRA KUMAR Principal Scientist, Animal Genetics Division, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 243 122 India
T K BHATTACHARYA National fellow, ICAR-DPR, Hyderabad. Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 243 122 India
A P SINGH Associate professor, Animal Biotechnology Centre, JNKVV Campus, Jabalpur
B BHUSHAN Principal Scientist, Animal Genetics Division, Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 243 122 India
A SHARMA Director, ICAR-NBAGR, Karnal. Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 243 122 India
A K TIWARI Head, Standardization Division. Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 243 122 India

https://doi.org/10.56093/ijans.v85i12.54366

Keywords:

Buffalo, cDNA sequencing, In-silico analysis, Macrophage lysozyme

Abstract

The role of immune-relevant genes involved in disease biology as well as innate and adaptive immunity is at the centre stage while exploring the genetic basis of disease occurrence. The macrophage expressed lysozyme gene product contributes to innate immunity by cleaving the ÃŸ- 1,4 linkage of bacterial peptidoglycan in phagocytosis. Higher specific activity observed in bubaline milk/ serum lysozymes over their bovine counterparts is expected to contribute higher resistances to diseases in general and mastitis in particular. To explore the reasons for higher activity in terms of sequence variations, macrophage expressed lysozyme cDNA was synthesized, cloned and sequenced. The 593 bp mRNA sequence revealed a 444 bp (27nt to 470nt) ORF bearing usual start codon ATG and end codon TAA with GC content of 46 % coding for precursor polypeptide of 147 amino acids. The comparative sequence analysis of cattle and buffalo revealed the difference at 10 places leading to 3 non-synonymous (5^th, 116^th and 142^th) substitutions which did not affect the predicted 3D structure. The mLYS polypeptide had signal sequence (1^st to 18^th residue) and the mature peptide had two lysozyme catalytic sites, three Ca++ binding sites as well as eleven catalytic clefts prominent among the conserved domains. Secondly, bubaline mLYS was phylogenetically closer to the abomasum type than mammary gland type. The high similarity in the coding sequences and predicted structure suggested that bubaline lysozyme gene if hyper-expressed either in native or recombinant form in bovine udder may result in better udder health.

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