Genotyping of major histocompatibility complex Class II DRB gene in Rohilkhandi goats by polymerase chain reaction-restriction fragment length polymorphism and DNA sequencing

Abstract

Aim:To study the major histocompatibility complex (MHC) Class II DRB1 gene polymorphism in Rohilkhandi goat using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and nucleotide sequencing techniques.Materials and Methods:DNA was isolated from 127 Rohilkhandi goats maintained at sheep and goat farm, Indian Veterinary Research Institute, Izatnagar, Bareilly. A 284 bp fragment of exon 2 of DRB1 gene was amplified and digested using BsaI and TaqI restriction enzymes. Population genetic parameters were calculated using Popgene v 1.32 and SAS 9.0. The genotypes were then sequenced using Sanger dideoxy chain termination method and were compared with related breeds/species using MEGA 6.0 and Megalign (DNASTAR) software.Results:TaqI locus showed three and BsaI locus showed two genotypes. Both the loci were found to be in Hardy–Weinberg equilibrium (HWE), however, population genetic parameters suggest that heterozygosity is still maintained in the population at both loci. Percent diversity and divergence matrix, as well as phylogenetic analysis revealed that the MHC Class II DRB1 gene of Rohilkhandi goats was found to be in close cluster with Garole and Scottish blackface sheep breeds as compared to other goat breeds included in the sequence comparison.Conclusion:The PCR-RFLP patterns showed population to be in HWE and absence of one genotype at one locus (BsaI), both the loci showed excess of one or the other homozygote genotype, however, effective number of alleles showed that allelic diversity is present in the population. Sequence comparison of DRB1 gene of Rohilkhandi goat with other sheep and goat breed assigned Rohilkhandi goat in divergence with Jamanupari and Angora goats.

A novel technique for retrospective genetic analysis of the response to vaccination or infection using cell-free DNA from archived sheep serum and plasma

“…The same PCR primer combinations amplified DRB1 from DNA isolated from the serum of Texel cross sheep and from bovine and caprine serum, demonstrating the wide applicability of the technique, although primer optimisation may be required for each species. In previous studies, the primer combinations employed in this study successfully amplified DRB1 from genomic DNA obtained from whole blood samples from a range of sheep breeds [6] and from goats [39,40]. In this study, amplification of the Ovar-DRB1 locus failed in 33 samples.…”

Dna Barcoding Confirma a Ocorrência De Espécies Amazônicas Na Ictiofauna Do Rio Turiaçu, Maranhão/Brasil

“…Embora o MHC seja considerado como uma região associada a resistência a doenças (YADAV et al, 2016), a caracterização completa dos genes que a compõem é difícil devido ao seu grande tamanho. Dessa forma, estudos têm sido realizados em fragmentos, de acordo com a importância de diferentes regiões (SHRIVASTAVA et al, 2015).…”

“…Amills et al (2004) relataram a existência de 22 alelos diferentes do locus DRB em cabras. No entanto, a pesquisa sobre a genética da resistência às doenças em cabras ainda é insipiente e pouco se sabe sobre a frequência alélica desse gene e as possíveis associações entre os seus alelos (PETLANE et al, 2012;SHRIVASTAVA et al, 2015). Além disso, esse gene tem sido relacionado com uma ampla variedade de características de produção, segurança e qualidade de produtos em animais domésticos (PARACHA et al, 2015).…”

Analysis of Genetic Variability in the Caprine Class II MHC CLA DRB3.2(Cahi DRB3.2) gene locus of the Sangamneri goat breed of India

“…The CLA-DRB3.2 encodes the beta-1 domain of the DR molecule, which is in close contact with the foreign antigen and displays a very high degree of polymorphism, with more than 25 different sequences identi ed to date (Schwaiger et al, 1993, Amills et al, 1995. Shrivastava et al, (2015), studied the genetic polymorphism present in the Class II MHC DRB gene in the Rohilkhandi goats of Western Uttar Pradesh by the technique of PCR-RFLP. Genetic diversity in the of the DQB1 locus of the Class II Major Histocompatibility Complex Class II in Nigerian goats was studied by Yakubu et al, 2013.…”