Mutations in CD14 gene causes mastitis in different breeds of buffalo as confirmed by in silico studies and experimental validation

Abstract

Background CD 14 is an important pattern recognition receptor having innate immune function and has antibacterial activity. It binds with LPS of gram-negative bacteria, arachidonic acid, and lipoteichoic acid. Being a receptor, it binds with the pathogen with the help of other cytokines. Mutations in CD14 affect the binding ability which in turn affects the biological potentiality. Method The present study was conducted on 228 nos. of buffaloes pertaining to four different breeds as Murrah, Mehsana, Surti and Bhadawari. CD14 gene was characterized and polymorphism was detected through Single nucleotide conformation polymorphism. Association study was conducted for different variants of CD14 with mastitis in buffalo, detected through somatic cell count, california mastitis test. Result Eight variants of CD14 were detected and mutational hotspots were detected in bubaline CD14 with 58 number of non-synonymous SNP, out of which 18 were observed to be deleterious and 34 as thermodynamically unstable. In the present study, we had detected the mutations in CD14 gene and its association with the somatic cell score and other indicators for mastitis. In-silico studies were conducted to understand the molecular mechanism how the mutations affect the biological potentiality by analyzing different domains and structural analysis along with various post-translational modification sites. Conclusion Deleterious mutations were observed in CD14 gene which have significant effect on mastitis of buffalo. It may be employed for marker assisted selection, therapeutic application of recombinant CD14, gene therapy, transgenic animal production with wild type CD14 resistant to mastitis as future strategy.

RIGI, TLR7, and TLR3 Genes Were Predicted to Have Immune Response Against Avian Influenza in Indigenous Ducks

“…The analysis of O-linked glycosylation sites was carried out using NetOGlyc 3.1 server ( http://www.expassy.org/ ), whereas the N-linked glycosylation site was detected by NetNGlyc 1.0 software ( http://www.expassy.org/ ). The detection of leucine-zipper was conducted through Expassy software, Technical University of Denmark ( Pal et al, 2020 ). Regions for alpha-helix and beta-sheet were predicted using NetSurfP-Protein Surface Accessibility and Secondary Structure Predictions, Technical University of Denmark ( Schlee et al, 2009 ; Pal et al, 2018a ).…”

“…expassy.org/), whereas the N-linked glycosylation site was detected by NetNGlyc 1.0 software (http://www.expassy.org/). The detection of leucine-zipper was conducted through Expassy software, Technical University of Denmark (Pal et al, 2020). Regions for alpha-helix and beta-sheet were predicted using NetSurfP-Protein Surface Accessibility and Secondary…”