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dc.contributor.authorBrown, Pamelaen
dc.date.accessioned2018-05-14T10:17:04Z
dc.date.available2018-05-14T10:17:04Z
dc.date.issued1990en
dc.identifier.urihttp://hdl.handle.net/1842/30048
dc.description.abstracten
dc.description.abstractThe bovine MHC class I loci have largely been characterised using immunological techniques. This has identified in excess of thirty alleles all corresponding to one locus, BoLA-A . Detailed analysis of bovine MHC class I genes was undertaken using recombinant DNA technology in an effort to expand on this knowledge.en
dc.description.abstractA bovine liver cDNA library was constructed in Xgt10 and screened with a human class I MHC DNA probe. Two bovine class I cDNA clones were isolated. The largest clone, pBoLA-1, size 1263bp, was incomplete lacking a transcription initiation codon and a polyadenylation signal. pBol_A-1 on translation into amino acid sequence could encode a mature protein of 339 amino acids. Comparison of the amino acid sequence of pBoLA-1 to two other bovine cDNA clones revealed that pBoLA-1 may be allelic to one of them. Analysis of these bovine clones facilitated the identification of putative species specific residues. Extensive comparison of pBoLA-1 protein to class I proteins of other species revealed that the protein was more similar to class la proteins than to class lb proteins. The second cDNA clone, pBoLA-2, was approximately 600bp in size. pBoLA-2 mapped to the 3' end of pBoLA-1, extending it to 1.7kb, the approximate size of class I transcripts. The exact nature of the sequence encoded by pBoLA-2 was not established, although Northern blots suggested it may contain a repeated element.en
dc.description.abstractTwo bovine genomic clones (which were isolated from a bovine genomic DNA library by Dr Jean-Luc Vilotte using the pBoLA-1 cDNA clone as a probe) were chosen for further characterisation. The first, phage 41, encoded two non-contiguous hybridising pieces of DNA. Restriction enzyme digests of this phage were compared on a Southern blot to similar digests of genomic DNA, the isolated phage clone appeared to contain a sequence which was probably present. as 1 copy in the genome. The conclusion reached was that phage 41 carries two class 1 genes, seperated by ISRb. This close proximity of class 1 genes has also been observed in mouse. The second genomic clone, phage 33, encoded a class I pseudogene. The gene lacked a defined 5' end but shared 52-76.5% nucleotide similarity with pBoLA-1 stretching over exons five to eight.en
dc.description.abstractLastly, pBoLA-1 was used to dissect the detailed multi-band hybridisation pattern obtained when probing Southern blots. Use of 5' and 3' probes allowed an estimate of at least 2-3 bands recognised in common. Use of a 5' probe at high stringency revealed that pBoLA-1 only detected polymorphic bands, these by inference are class la genes with nonpolymorphic bands corresponding to class lb genes. The polymorphic bands recognised by pBoLA-1 were correlated to the BoLA-A serological type of the animal. Bands which segregated with BoLA-w6.2, -w8 an6-w10 were identified.en
dc.description.abstractKnowledge of the class I genes of the bovine MHC was extended from a single serologically defined locus to a large multi-gene family. This multi-gene family consists of class la genes which include pBoLA-1 and possibly class lb genes as defined by genomic clones and Southern blotting analysis of bovine DNA.en
dc.publisherThe University of Edinburghen
dc.relation.ispartofAnnexe Thesis Digitisation Project 2018 Block 18en
dc.relation.isreferencedbyAlready catalogueden
dc.titleCloning and characterisation of MHC class 1 genes in cattleen
dc.typeThesis or Dissertationen
dc.type.qualificationlevelDoctoralen
dc.type.qualificationnamePhD Doctor of Philosophyen


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