Bovine blood group genes : somatic cell mapping and homologous relationships with the human systems RH and ABO /
Primer pairs designed for the amplification by polymerase chain reaction (PCR) of the genes encoding the human blood group systems Rh and ABO were used on genomic DNA from human and bovine to produce probes containing segments of this genes. Two different set of primers were used for the Rh system,...
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| Format: | Thesis Book |
| Language: | English |
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[Place of publication not identified] :
[publisher not identified] ;
1995.
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| Online Access: | http://proxy.library.tamu.edu/login?url=http://proquest.umi.com/pqdweb?did=742535301&sid=1&Fmt=2&clientId=2945&RQT=309&VName=PQD |
| Summary: | Primer pairs designed for the amplification by polymerase chain reaction (PCR) of the genes encoding the human blood group systems Rh and ABO were used on genomic DNA from human and bovine to produce probes containing segments of this genes. Two different set of primers were used for the Rh system, one for the RhD gene and other for the RHCE gene. The primers for the ABO system were designed to amplifiy the A gene (A-transferase). Additionally, the primers were used on genomic DNA from bovine-rodent somatic cell hybrids segregating bovine chromosomes to determine the chromosomic localization of the Rh and ABO genes in the bovine genome. PCR on bovine DNA amplified a 950 bp long product with primers for the RHCE gene in some animals but not in others. Four bovine B blood group system factors (01, P, O', and I') were found to be present only in PCR-positive animals. Amplification of bovine DNA with primers for the RhD gene yielded no detectable product, confirming the ancestral nature of the RHCE gene.Southern blotting of human and bovinea probes containing the RHCE gene to genomic bovine DNA produced similar hybridization patterns and detected four restriction fragments in some animals and three in others. A relationship between intensity of the hybridization signal and the phenotype of the bovine blood group B was detected; animals with phenotypes containing factors present only in the PCR-positive animals (01 and O') presented stronger signals. This result suggests an homologous relationship among human RH and bovine B blood groups, supporting reported serological and biochemical similarities. A moderate nucleotide sequence conservation (69%) between the bovine RHCE probes and human Rh was found, indicating evolutionary divergence. PCR-somatic cell mapping of this bovine probe assigned bovine Rh to chromosome 3, further defining a region of conserved synteny with human chromosome 1. Primers for the amplification of human gene encoding the A allele of the ABO blood group (A-transferase) produced a 350 bp long product in bovine DNA. No correlation was found between blood type and presence of amplification, and several hybridization bands were detected by Southern blotting of human probes of the A allele to bovine DNA. Nucleotide sequence comparison among bovine A-transferase clones revealed a low conservation (52%) with the human gene. Physical mapping of the bovine probe yielded a tentative assignment (88 % concordance) to chromosome 8, further defining a region of conserved synteny with human chromosome 9. The ability to amplify specific sequences on bovine DNA with PCR primers for human systems Rh and ABO, as well as the Southern blotting experiments indicate the presence of related sequences in the bovine genome. However, these sequences may represent non-coding or ancestral genes, since the degree of nucleotide sequence conservation was either low (A gene) or moderate (RhC gene), and because the sequences amplified in this study were not mapped to chromosomes on which the possible homologous of the human system have been located. |
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| Item Description: | Vita. "Major Subject: Veterinary Microbiology". |
| Physical Description: | xi, 83 leaves : illustrations ; 28 cm. Issued also on microfiche from University Microfilms Inc. |
| Bibliography: | Includes bibliographical references. |