Biomaterial Database

A PCR-RFLP typing method for the caprine Mhc class II DRB gene. 1996

M Amills, and O Francino, and A Sànchez

Departament de Patologia i Producció Animals, Facultat de Veterinària, Edifici V, Universitat Autònoma de Barcelona, Bellaterra Spain. marcel@guara.uab.es

A polymerase chain reaction (PCR) technique associated with restriction fragment length polymorphism (RFLP) analysis has been developed for typing the second exon of the caprine Mhc class II DRB gene. By using a maximum of three restriction enzymes, corresponding to different combinations of RsaI, BsaI, AccI, NdeII, HaeIII and HpaII, this procedure allows to distinguish unequivocally 18 of the 22 caprine DRB alleles. Forty goats of several breeds were typed with this technique and two new RsaI restriction patterns which did not match with previously published sequence data were identified. Close associations have been found between RFLPs and amino acid substitutions at positions which are expected to be involved in the formation of the antigen-recognition site (ARS) of the DR molecule. These results suggest that PCR-RFLP may be a useful tool in typing the caprine DRB gene and in relating amino acid substitutions at the ARS of the DR molecule with disease resistance.

UI	MeSH Term	Description	Entries
D012150	Polymorphism, Restriction Fragment Length	Variation occurring within a species in the presence or length of DNA fragment generated by a specific endonuclease at a specific site in the genome. Such variations are generated by mutations that create or abolish recognition sites for these enzymes or change the length of the fragment.	RFLP,Restriction Fragment Length Polymorphism,RFLPs,Restriction Fragment Length Polymorphisms
D005802	Genes, MHC Class II	Genetic loci in the vertebrate major histocompatibility complex that encode polymorphic products which control the immune response to specific antigens. The genes are found in the HLA-D region in humans and include H-2M, I-A, and I-E loci in mice.	Class II Genes,Genes, Class II,Genes, HLA Class II,MHC Class II Genes,Class II Gene,Gene, Class II
D006041	Goats	Any of numerous agile, hollow-horned RUMINANTS of the genus Capra, in the family Bovidae, closely related to the SHEEP.	Capra,Capras,Goat
D000483	Alleles	Variant forms of the same gene, occupying the same locus on homologous CHROMOSOMES, and governing the variants in production of the same gene product.	Allelomorphs,Allele,Allelomorph
D000818	Animals	Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA.	Animal,Metazoa,Animalia
D016133	Polymerase Chain Reaction	In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships.	Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain