Technical Note: DNA Typing for Ovine MHC DRB1 Using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP)

Abstract

The ovine major histocompatibilty complex (Ovar) class II DRB1 second exon was amplified by polymerase chain reaction (PCR) from DNA samples of 52 Suffolk sheep. Polymerase chain reaction products were characterized by the restriction fragment length polymorphism (RFLP) technique using nine restriction enzymes, RsaI, HaeIII, SacI, SacII, DdeI, NciI, Hin1I, EcoRI, and BstNI, yielding 13 types. Sequencing of cloned PCR products identified 16 Ovar-DRB1 alleles. Collectively, all PCR-RFLP patterns exactly matched those predicted from DNA sequences. These findings strongly indicate that the PCR-RFLP method using a combination of nine restriction endonucleases is a very powerful tool in Ovar typing.

Identification of genetic variation in the major histocompatibility complex gene region in Turkish sheep breeds

“…Alleles found in this study were similar to those previously identified and reported by Konnai et al (2003) and Amills et al (1996). Although novel alleles were not identified, further detailed DNA sequence analysis of the exon 2 regions of Ovar-DRB1 and Ovar-DRB3 from Turkish sheep breeds will be carried out.…”

“…Although novel alleles were not identified, further detailed DNA sequence analysis of the exon 2 regions of Ovar-DRB1 and Ovar-DRB3 from Turkish sheep breeds will be carried out. (Amills et al 1996;Konnai et al 2003;Dongxiao & Yuan 2004;Gruszczynska et al 2005;Brujeni et al 2009). To date, 106 Ovar-DRB1 alleles have been identified by DNA sequencing of exon 2 from various breeds of sheep (Schwaiger et al, 1995;Schwaiger et al, 1996;Jugo & Vicario, 2000;Konnai et al, 2003;Ballingall et al, 2008).…”

“…The MHC class III region encodes other immune components including the complement system (e.g., C2, C4 or factor B) and cytokines (e.g., TNF-α). Studies of genetic variation in Ovar-Mhc class II genes have shown that the expressed DRB locus is highly polymorphic (Schwaiger & Epplen, 1995;Schwaiger et al, 1996;Jugo & Vicario, 2000;Konnai et al, 2003;Ballingall et al, 2008;Nikbakht et al, 2012;Lotfi et al, 2012;Shen et al, 2014;Takeshima et al, 2014). In particular, a high polymorphism level is present in exon 2, which encodes the antigen-binding site (Escayg et al, 1997;Konnai et al, 2003).…”

“…OLA-DRB1 was amplified according to Konnai et al (2003). Nested PCR was used to amplify the second exon of the DRB1 gene.…”

Association of molecular polymorphism in exon 2 of Ovar-DRB1 gene with weight traits in Iranian Makuie sheep breed

“…The MHC of sheep is known as Ovar ('Ovar' representing Ovis aries) and is located on chromosome 20. The Ovar class II genes encode polymorphic glycoproteins composed of 9 covalently linked subunits (α and β subunits) that play a pivotal role in the initiation of the immune response against pathogen-derived peptide antigens (11). Among Ovar class II genes, the expressed DRB1 locus has been found to be highly polymorphic, and most of the polymorphic sites on DRB1 are located in exon 2, which encode the outer domain of MHC, especially the β-chain, which is the binding site for an antigen (11,12).…”