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Biomedical subjects

S J Kemp

Publications and source records attributed to S J Kemp.

At least 19 recordsLinked to original sources

A medium-density genetic linkage map of the bovine genome.

A cattle genetic linkage map was constructed which covers more than 95 percent of the bovine genome at medium density. Seven hundred and forty six DNA polymorphisms were genotyped in cattle families which comprise 347 individuals in full sibling pedigrees. Seven hundred and three of the loci are linked to at least one other locus. All linkage groups are assigned to chromosomes, and all are orientated with regards to the centromere. There is little overall difference in the lengths of the bull and cow linkage maps although there are individual differences between maps of chromosomes. One hundred and sixty polymorphisms are in or near genes, and the resultant genome-wide comparative analyses indicate that while there is greater conservation of synteny between cattle and humans compared with mice, the conservation of gene order between cattle and humans is much less than would be expected from the conservation of synteny. This map provides a basis for high-resolution mapping of the bovine genome with physical resources such as Yeast and Bacterial Artificial Chromosomes as well as providing the underpinning for the interpolation of information from the Human Genome Project.

Animals

Sensitivity of segregation analysis to data structure and transformation: a case study of trypanotolerance in mice.

Sensitivity of segregation analysis for data structure and data transformation was studied using data from two trials in which mice were challenged at three months of age with a cloned isolate of Trypanosoma congolense and survival time was recorded. Data included records from three inbred strains (C57BL/6 (tolerant), A/J, and BALB/c (both susceptible)) and their crosses. Data were standardized and normalized using a modified power transformation. Segregation analysis was applied to both untransformed and transformed data to determine the genetic inheritance of trypanotolerance in these mice. Data from the two trials were analysed separately and combined. Four genetic models were compared; a one locus model, a polygenic model, a mixed model with common variance, and a mixed model with different variances for each major genotype. Even though the separate data sets and the combined data set all supported the hypothesis of a major gene (or a tightly linked cluster of genes) with different variances within each genotype, parameter estimates were highly sensitive to data transformation and several sets of parameter estimates gave similar likelihood values because of high dependency between parameters. Based on the results segregation analysis can be very sensitive to data structure in a crossbreeding design and to data transformation. Interpretation of the results can be misleading if the entire parameter space is not studied carefully.

Animals

Localization of genes controlling resistance to trypanosomiasis in mice.

Tsetse fly-transmitted trypanosomes (Trypanosoma spp.) cause "sleeping sickness' in man and have a serious impact on livestock-based agriculture in large areas of Africa. Multigene control of variation in susceptibility to trypanosomiasis is known to occur in mice, where the C57BI/6 (B6) strain is relatively resistant and the A/J (A) and Balb/c (B) strains are susceptible. Such resistance is also well described among several types of west African cattle. We report here the results of genome-wide scans for genes controlling this trait in the B6 mouse using crosses with two different susceptible strains. Regions on mouse chromosomes 5 and 17 were found to be important in determining resistance in both crosses while an additional region on chromosome 1 showed evidence of involvement in only one cross. We confirmed the size of the effect due to chromosome 17 in F3 intercross populations fixed for alternative parental chromosomes. The three loci are of large effect and account for most of the genetic variation in both F2 populations. We propose that they be designated Tir1, Tir2 and Tir3.

Animals

Involvement of protein tyrosine kinases in regulation of endothelial cell organization by basement membrane proteins.

During capillary formation, endothelial cell migration and organization are critically dependent on surrounding basement membrane proteins. These proteins serve as a physical support and are likely to provide signals which regulate migration and organization of the cells. In this study the possible involvement of tyrosine kinase signalling pathways in basement membrane-induced organization of human endothelial cells is examined. Interaction of endothelial cells with reconstituted basement membrane Matrigel activates tyrosine phosphorylation of several proteins including focal adhesion kinase. Inhibition of this pathway with tyrosine kinase inhibitors impairs localization of paxillin to focal adhesions and organization of actin filaments, decreases motility and elongation of endothelial cells, and prevents their organization into cords or tubes on basement membrane. These data demonstrate that basement membrane-induced modulation of endothelial cell motility, shape, and organization is critically dependent on tyrosine kinase signalling pathway(s) involving cytoskeletal proteins.

Basement Membrane

Polymorphism at the bovine tumor necrosis factor alpha locus and assignment to BTA 23.

We have identified four single-strand conformation variants of the bovine tumor necrosis factor alpha gene by analysis of PCR-amplified fragments. The variants are inherited in Mendelian fashion and are informative for linkage mapping. We have mapped the bovine gene to Chromosome (Chr) 23 in a panel of somatic cell hybrids and observed genetic linkage to the major histocompatibility complex (BoLA) genes and microsatellite markers on bovine Chr 23 in an international bovine reference family panel. The distribution of the alleles was determined in cattle of different breeds and of different geographical origins, which included trypano-susceptible and trypano-tolerant cattle.

Animals

Genetic control of resistance to trypanosomiasis.

To map the genetic sources of trypanotolerance in mice, a linkage analysis of survival following trypanosome challenge was performed by selective genotyping in a large F2 population produced by crossing the resistant C57BL/6 and susceptible BALB/c inbred mouse lines. We report evidence of a chromosomal region of large effect, possibly comprising more than one resistance locus, on Chromosome 17; and of further loci on Chromosomes 1 and 5. Together, these genes can account for all of the difference between the mean parental phenotypes.

Animals

Parasite strain specificity of bovine cytotoxic T cell responses to Theileria parva is determined primarily by immunodominance.

The parasite strain specificity of CTL responses to Theileria parva varies among cattle immunized with the same parasite stock. We have investigated the influence of class I MHC on the strain specificity of CTL responses to T. parva in 19 cattle of defined class I phenotype immunized with either of two T. parva populations, in which protection to subsequent reciprocal challenge correlated with CTL strain specificity. In the majority of animals the response was restricted by the products of one MHC haplotype and there was a consistent bias to some haplotypes in preference to others. In 10 of 13 cattle expressing the molecularly defined MHC specificities A10 and KN104 on one haplotype, the CTL response was restricted entirely by this haplotype, thus allowing a precise analysis of the MHC restriction specificities. The MHC restriction specificity and the parasite population used for immunization both influenced the strain specificity of the response. By examining responses in identical twins immunized with different parasites or in animals before and after challenge with heterologous parasites, animals that mounted a strain-specific response to primary infection were shown to be capable of responding to Ags shared by the two parasite populations. These findings indicate that the strain specificity of CTL responses to T. parva is not determined primarily by immune response genes that define the inherent capacity to respond, but rather is a consequence of the response in individual animals being biased toward a limited number of immunodominant peptide-MHC determinants.

Animals

Sesamoid arthrodesis for hyperextension of the thumb metacarpophalangeal joint.

Forty-two sesamoid arthrodeses performed since 1986 were reviewed. Thirty-seven cases were considered successful. Hypertension recurred in 3 of the 20 procedures performed for cerebral palsy and 1 of the 21 performed in association with basal joint arthroplasty for arthritic conditions. The single post trauma case was successful. Flexion was preserved. Prevention of hyperextension of the metacarpophalangeal joint assists in reducing metacarpal adduction in cerebral palsy and has a stabilizing effect in basal joint arthroplasty.

Adolescent

A panel of polymorphic bovine, ovine and caprine microsatellite markers.

A panel of 81 new polymorphic bovine microsatellite markers is described, together with further information on a previously reported group of 16 markers. The mean polymorphism information content of the 97 markers determined in 20 cattle was 0.66. Seventy-three of these markers have been assigned to chromosomes by either linkage analysis or use of hybrid cell panels. Thirty-nine of the markers were polymorphic in sheep, and 32 were polymorphic in goat. This study identified a set of 18 robust markers that were polymorphic in all three species and that covered 14 bovine chromosomes. This provides a single group of markers, which would be suited to genetic distance analysis and parentage control in cattle, sheep and goat.

Animals

A polymorphism in randomly amplified DNA that differentiates the Y chromosomes of Bos indicus and Bos taurus.

A small number of west African Bos taurus cattle breeds, including the N'Dama, constitute a valuable genetic resource by virtue of their ability to remain productive under trypanosomiasis challenge. However, introgression of Bos indicus genes into the trypanotolerant breeds, particularly by introduction of zebu bulls, is a threat to this resource. This work describes the characterization and cloning of a bovine randomly amplified polymorphic DNA (RAPD) that is generated in polymorphic DNA (RAPD) that is generated in polymerase chain reaction (PCR) with the 10 base primer ILO1065 from Bos indicus male templates, but not from B. taurus male templates or female templates of either type. Male-specific sequences with homology to the RAPD also occur in B. taurus breeds. This suggests that the polymorphism may be due to base substitution(s) in an ILO1065 priming site, or insertion/deletion events either affecting priming sites or occurring between sites on the cattle Y chromosome. We have shown that cattle, whether of B. indicus or B. taurus phenotype, which possess a typically B. indicus metaphase Y chromosome on the basis of QFQ banding, have a B. indicus ILO1065-generated genotype. The ILO1065-primed RAPD can be used in a simple dot blot assay as a probe of RAPD-PCR products, to provide a convenient, reliable and effective means of detecting introgression of zebu genes in B. taurus cattle populations.

Africa, Western

Fifteen new synteny assignments of microsatellites to the bovine genome.

A panel of 36 hamster-bovine hybrid cell lines was used to assign 15 bovine microsatellites. Locus identification, synteny group and/or chromosome assignment and registration number were as follows: ILSTS001 (U22, Chr07, D7S13), ILSTS002 (U09, Chr18, D18S7), ILSTS004 (U10, Chr01, D1S28), ILSTS005 (U05, Chr10, D10S25), ILSTS006 (U22, Chr07, D7S8), ILSTS008 (U24, Chr14, D14S15), ILSTS010 (U27, DU27S11), ILSTS011 (U24, Chr14, D14S16), ILSTS012 (U16, Chr11, D11S26), ILSTS015 (U07, Chr25, D25S3), ILSTS016 (U04, Chr21, D21S22), ILSTS017 (X, DXS11), ILSTS018 (U15, Chr06, D6S15), ILSTS019 (U07, Chr25, D25S7) and ILSTS020 (U05, Chr10, D10S27). These results contribute to the international effort to improve the bovine genetic map.

Animals

PCR-RFLP typing of the bovine myoglobin gene.

We have identified substitutions in the 3' untranslated region of the bovine myoglobin gene, one of which affects an MboII restriction enzyme site resulting in a bi-allelic restriction fragment length polymorphism. Co-dominant inheritance of the alleles in three reference families was observed using a polymerase chain reaction--restriction fragment length polymorphism assay. The distribution of the alleles seems characteristic of cattle type--one of the alleles was not detected in purely taurine breeds. Furthermore, we mapped, using the polymerase chain reaction on a bovine-rodent somatic cell hybrid panel, the myoglobin gene to bovine chromosome five. It is therefore syntenic with gamma-interferon and insulin-like growth factor in which we have not found polymorphism. The myoglobin locus therefore serves as a type one marker on bovine chromosome five.

Alleles

Randomly primed PCR amplification of pooled DNA reveals polymorphism in a ruminant repetitive DNA sequence which differentiates Bos indicus and B. taurus.

By amplification of pools of DNA representative of different bovine populations with single short oligonucleotide primers of random sequence, we were able rapidly to identify markers which distinguish the two major subspecies of domestic cattle, Bos taurus and B. indicus. One of the marker polymorphisms was found to be in a novel, dispersed DNA sequence which occurs in several ruminant species. The marker will assist in the detection of crossbreeding between Zebu and B. taurus types where this threatens a potentially valuable trypanosomiasis-resistant B. taurus genetic resource in West Africa. In addition, the marker will be useful for exploration of the evolutionary relationships of the major subspecies of domestic cattle. The general approach used to identify population-specific DNA polymorphisms has potentially broad application in definition of species, breeds and populations and will be of generic value in studies of genome evolution.

Animals

Characterization of Zebu cattle breeds in Tanzania using random amplified polymorphic DNA markers.

A total of 141 short primers, of arbitrary nucleotide sequence, were used singly in polymerase chain reactions to amplify DNA fingerprints in pools of DNA representing three Zebu cattle breeds. Two primers, which discriminated between the breed-specific DNA pools were used further to amplify individual pool components in order to establish band frequencies of the amplified fingerprints. One of the primers (ILO 1127) amplified a RAPD fingerprint in 61% of TSZ animals but less than 6% in the other breeds, while another primer (ILO 1065) revealed a DNA sequence common to 89% of the Boran animals and less than 30% in the other two breeds. Bandsharing and mean average percentage difference calculated within and between the three breeds using RAPD fingerprint data showed a higher degree of homogeneity within than across the breeds and indicated measurable divergence between the three breeds. It is concluded that RAPD polymorphisms are useful as genetic markers for cattle breed differentiation.

Animals

A polymorphism in the bovine gamma-S-crystallin gene revealed by allele-specific amplification.

A polymorphism was detected in the 3' untranslated region of the bovine gamma-S-crystallin gene by direct sequencing of polymerase chain reaction (PCR) products from genomic DNA of an N'Dama bull and a Boran cow. A set of three PCR primers was designed to detect this difference and thus give allele-specific amplification. The two allele-specific primers differ in length by 20 nucleotides so that the allelic products may be distinguished by simple agarose gel electrophoresis following a single PCR reaction. This provides a simple and rapid assay for this polymorphism.

Alleles