Restriction fragment alleles of the rabbit IGHG genes with reference to the rabbit IGHGCH2 or e locus polymorphism

Abstract

Among domesticated mammals, rabbit (Oryctolagus cuniculus) is the only species possessing not more than one subclass of immunoglobulin (IgG) antibodies. The rabbit IGHGCH2 or e locus presents two serologically defined alleles, the e14 and e15 allotypes, which are correlated with amino acid variation at the IgG CH2-CH3 interface. Genetic studies, while revealing the adaptive value of this polymorphism, have relied so far entirely upon allo-antisera. Here we show how these alleles can be distinguished by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods. The proposed PCR-RFLP approach allows the monitoring of IGHG locus diversity in rabbit.

Detection of RHDV strains in the Iberian hare ( Lepus granatensis ): earliest evidence of rabbit lagovirus cross-species infection

“…Total genomic DNA was extracted from the liver samples using the EasySpin Genomic DNA Minipreps Tissue Kit (Citomed, Lisbon, Portugal) according to the manufacturer's instructions. The nuclear markers have been shown to present specific SNP for Oryctolagus and Lepus and their amplification followed the conditions described in [14][15][16]. The amplification of the mitochondrial marker was performed using the primers described in [17].…”

Molecular bases of genetic diversity and evolution of the immunoglobulin heavy chain variable region (IGHV) gene locus in leporids

“…The study of these species has yielded some interesting results and led to new insights on the evolution of IGHV genes in the Lagomorph group. Also, the few data obtained so far for the Lagomorph IGHG hinge region and CH2 domain has shown some particular patterns of genetic diversity possibly linked to resistance against pathogens (Esteves et al, 2002a; Esteves et al, 2002b; Esteves et al, 2006). Extending our knowledge about both the IGHV genes and IGH constant regions to other Lagomorph species would certainly contribute further to our understanding of the evolution of IGHV genes and the roles that genetic diversity of immunoglobulins may play in host resistance to pathogens.…”

Genome Mapping and Genomics in Domestic Animals

“…The rabbit genome carries only one immunoglobulin gamma class. DNA typing methods based on sequencing and PCR-RFLP techniques have recently been set up to upgrade the serological methods for allele identification of IGH genes, thus providing additional typing tools in the absence of antisera (Esteves et al 2002(Esteves et al , 2004(Esteves et al , 2006. The d locus corresponds to the hinge H and the e locus to the CH2 exon encoding a C-domain (Esteves et al 2006).…”

“…The d locus corresponds to the hinge H and the e locus to the CH2 exon encoding a C-domain (Esteves et al 2006). The PCR-RFLP typing designed for this locus consists in the amplification of a 553-557 bp fragment spanning the CH2 exon followed by a ThaI restriction enzyme digestion (Esteves et al 2002). At the e locus, the e14 and e15 allotypes have been defined, which correlate with an alanine-threonine exchange at amino acid position 309 (Appella et al 1971;Lefranc et al 2005).…”