Comparative molecular characterization and In silico analysis of neck and carbohydrate recognition domain (NCRD) of conglutin in gene in buffalo (Bubalis bubalis) and nilgai (Boselaphus tragocamelus)

Abstract

Conglutinin, a high molecular-weight C-type lectin belongs to the family of collectins, was originally detected in bovine serum which binds sugar residues in a Ca2+-dependent manner and it is an effector molecules in innate immunity. Total RNA extracted from liver sample of buffalo and nilgai were used to synthesize cDNA. Conglutinin NCR domain cDNA (497 bp) was amplified by proof reading DNA polymerase using primers established for cattle (Bostaurus). The amplicons were ligated to cloning vector (pJET), that was transformed into E. coli DH5α competent cells and the recombinant plasmids (pJET-BuCGN and pJET-NCGN) were characterized by Pst I restriction analysis. The amplicon (497 bp) was released from the recombinant plasmids (pJET-BuCGN and pJET-NCGN) upon Eco RI and Xho I double digestion, sequencing of the recombinant plasmids were carried out. BuCGN was found to be 99.6% similar to NCGN at nucleotde level, and 100% at predicted amino acid level. The predicted proteins of both the species (162 amino acids) were expected to possess five β-sheets in the secondary structure.