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Biomedical subjects

B W Kirkpatrick

Publications and source records attributed to B W Kirkpatrick.

At least 19 recordsLinked to original sources

Comparative linkage mapping of human chromosome 13 and bovine chromosome 12.

A comparative linkage map of human chromosome 13 and bovine chromosome 12 was constructed using eight polymorphic microsatellite markers associated with six specific genes. Linkage of these was also examined relative to five previously mapped anonymous microsatellite markers. Seven gene-linked markers were developed from bovine large-insert genomic clones containing one of five genes of interest (serotonin receptor subtype 2, fms-related tyrosine kinase, coagulation factor 10, retinoblastoma susceptibility gene, collagen type IV alpha 1), and one additional marker was developed from a microsatellite resident within an intron of the bovine dopachrome tautomerase gene. Four of these loci were previously assigned to bovine chromosome 12 by analysis of a somatic cell hybrid panel. This study provides linkage information for examining gene order in this conserved synteny group. The comparative linkage mapping results indicate that the q arm of human chromosome 13 is almost entirely conserved in bovine chromosome 12. One intrachromosomal rearrangement was detected in this linkage group relative to human, and this rearrangement was confirmed by fluorescence in situ hybridization results.

Animals

Relationship of growth hormone and insulin-like growth factor-1 genotypes with growth and carcass traits in swine.

The contribution of chromosomal regions linked to growth hormone (GH) and insulin-like growth factor-1 (IGF-1) loci to variation in preweaning average daily gain, postweaning average daily gain (ADG), 10th rib backfat, loin-eye area and muscle pH were evaluated. Offspring of four purebred sires (A-D; n = 150, 195, 148 and 136, respectively) and two cross-bred sires (E and F; n = 157 and 145, respectively) were genotyped initially with GH and IGF-1 markers. When results of single marker analysis suggested possible linkage with a quantitative trait locus (QTL), additional flanking markers were typed for the family and interval mapping was performed. Growth hormone genotype was not associated with the traits evaluated in the study. Evidence suggestive of linkage was found for IGF-1 genotype and ADG in one sire family (lod = 2.3) where differences were 0.032 +/- 0.01 kg/day for alternative sire alleles. Evidence for a putative ADG QTL was greatest in the interval between IGF-1 and Sw1071. A similar genomic region has been associated with growth variation in mice; however, QTL mapping precision in the current study is insufficient to establish similarity.

Alleles

Mapping genomic regions associated with growth rate in pigs.

The objective of this study was to identify genomic regions associated with postweaning average daily gain (ADG) in swine. Two families were produced using sires obtained from the cross of lines divergently selected for high and low growth rate. Sires were mated to 29 unrelated dams that produced 124 and 115 offspring. Average daily gain was recorded for every individual; mean ADG was .593 +/- .007 kg/d and .619 +/- .009 kg/d in the two families. Selective genotyping was performed as a prescreening procedure to identify genomic regions potentially associated with ADG. A total of 75 genetic markers were used between both families. Markers identified as nominally significant (P < .05) in an individual marker analysis were subsequently typed in all individuals. Markers still nominally significant (P < .05) after analysis with the complete family were then reevaluated together with flanking markers in an interval mapping analysis. A single region on chromosome 3, flanked by markers Sw2429 and Sw251, for the second family had a logarithm of the odds score of 2.9, which is suggestive of linkage with a quantitative trait locus for ADG. Differences were .033 +/- .009 kg/d between individuals inheriting alternative paternal alleles for Sw251, which was the most significant marker.

Alleles

Exploiting dinucleotide microsatellites conserved among mammalian species.

Dinucleotide microsatellites are useful for gene mapping projects. Depending upon definition of conservation, published estimates of dinucleotide microsatellite conservation levels vary dramatically (30% to 100%). This study focused on well-characterized genes that contain microsatellites in the human genome. The objective was to examine the feasibility of developing microsatellite markers within genes on the basis of the assumption of microsatellite conservation across distantly related species. Eight genes (Gamma-actin, carcinoembryonic antigen, apolipoprotein A-II, cardiac beta myosin heavy chain, laminin B2 chain, MHC class I CD8 alpha chain, c-reactive protein, and retinoblastoma susceptibility protein) containing large dinucleotide repeat units (N > or = 15), complete genomic structure information, and homologous gene sequences in a second species were selected. Heterologous primers were designed from conserved exon sequences flanking a microsatellite motif. PCR products from bovine and porcine genomic DNA were tested for the presence of microsatellite sequences by Southern blot hybridization with biotin-labeled (CA)12 oligonucleotides. Fragments containing microsatellites were cloned and sequenced. Homology was verified by sequence comparisons between human and corresponding bovine or porcine fragments. Four of sixteen (25%) cross-amplified PCR products contained dinucleotide repetitive sequences with repeat unit lengths of 5 to 23. Two dinucleotide repetitive sequences showed microsatellite length polymorphism, and an additional sequence displayed single-strand conformational polymorphism. Results from this study suggest that exploitation of conserved microsatellite sequences is a useful approach for developing specific genetic markers for comparative mapping purposes.

Animals

Syntenic assignment of human serotonin receptor subtype 2 (HTR2), esterase D (ESD), and fms-related tyrosine kinase (FLT) homologs to bovine chromosome 12.

Bovine x rodent somatic hybrid cells have been used to syntenically map three bovine genes homologous to loci on human Chromosome (Chr) 13. These three loci, fms-related tyrosine kinase gene (FLT), esterase D (ESD), and 5-hydroxytryptamine receptor 2 (HTR2; serotonin receptor subtype 2), were assigned to bovine Chr 12 (BTA12) with 91-95% concordance to the coagulation factor 10 (F10) locus. Along with a previously mapped BTA12 gene, retinoblastoma-1 (RB1), this conserved synteny group spans 178 cM on human Chr 13 (HSA13). Previous reports suggested homology between HSA13 and both BTA2 and BTA12. Results reported here extend the boundary of the HSA13-BTA12 comparative map, contradict the previous preliminary assignment of ESD to BTA2, and suggest instead that the q arm of HSA13 may be entirely conserved in BTA12.

Animals

Report of the first workshop on the genetic map of bovine chromosome 23.

A report of the first workshop on the genetic map of bovine chromosome 23 (BTA23) is given. Five laboratories contributed data from 29 loci, including a total 11586 informative genotypes. The combined pedigrees represented 1930 potentially informative meioses. Eighteen of the 29 loci were common to two or more data sets and were used to construct a framework linkage map of BTA23. Twelve of the 18 could be ordered on the linkage map with a likelihood ratio of greater than 1000:1. Thus, a low resolution consensus map was constructed with a high level of support for order. The sex-averaged, female and male maps span 54.5, 52.7 and 55.8 cM, respectively. Sex-specific differences in recombination frequency were identified for eight pairs of framework loci. Average genetic distance between framework loci on the sex-averaged map is 5.0 cM.

Animals

Syntenic assignment of dopamine tautomerase (DCT) to bovine chromosome 12.

Heterologous primers were used to amplify an exon and intron-containing segment of the bovine homologue of the human dopachrome tautomerase gene. After confirmation of homology by sequence analysis (exon sequence similarity greater than 90%), bovine-specific primers were developed for synteny mapping purposes. The dopachrome tautomerase gene was assigned to bovine chromosome 12 (BTA12) with 97% concordance to the coagulation factor 10 locus. Together with previous synteny mapping of bovine chromosome 12 genes, fms-related tyrosine kinase, esterase D and 5-hydroxytryptamine receptor 2, this assignment further indicates conservation between human chromosome 13q and bovine chromosome 12.

Animals

A search for quantitative trait loci for ovulation rate in cattle.

Seventy-seven polymorphic microsatellites were analysed in offspring of three elite sires that were part of the foundation of an experimental population selected for twinning rate at the US Meat Animal Research Center, Clay Center, Nebraska. All females were assessed for ovulation rate by rectal palpation of corpora lutea over 8-10 consecutive oestrous cycles from approximately 12 to 18 months of age, and associations between ovulation rate and sire allele were examined in each of the three sire groups. A preliminary analysis was performed using selectively genotyped daughters of each sire. Markers found significant or approaching significance were also genotyped in all daughters, sons and granddaughters of these sires. A test of marker associations limited to the granddaughter data provided an independent confirmation of marker effect and significance relative to the initial test with daughter data. Putative ovulation rate quantitative trait loci were detected on chromosomes 7 and 23. Marker UWCA20 on chromosome 7 was associated with an effect in excess of one phenotypic standard deviation and accounted for approximately 10% of phenotypic variation ovulation rate. Marker CYP21 (steroid 21-hydroxylase) on chromosome 23 was associated with an effect of slightly less than half a phenotypic standard deviation and accounted for approximately 4% of phenotypic variation.

Alleles

Seven genes from human chromosome 18 map to chromosome 24 in the bovine.

Bovine sequence tagged sites (STSs) were developed for seven genes and used for synteny mapping with a hybrid bovine x rodent cell line panel. The genes were thymidylate synthase (TYMS), pituitary adenylate cyclase activating peptide (ADCYAP1), and melanocortin-2 receptor (MC2R) from the short arm of human chromosome (HSA) 18 and N-cadherin (CDH2), transthyretin (TTR), gastrin-releasing peptide (GRP), and plasminogen activator inhibitor 2 (PAI2) from the long arm of HSA 18. Primers for these genes were designed with human, ovine, or bovine sequences aligned with a sequence from a second species. The bovine PCR product was cloned, and the fragment was sequenced to verify that the homologous gene was indeed amplified. A second set of bovine-specific PCR primers were developed for each gene from these sequences. These STSs were used for synteny mapping, and all seven genes were syntenic with markers of bovine chromosome (BTA) 24. The concordance with BTA 24 was at least 96.5% for all genes.

Animals

Evaluation of different amplification protocols for use in primer-extension preamplification.

Different amplification protocols were evaluated for use with primer-extension preamplification (PEP). We hypothesized that a protocol known to improve amplification of long DNA fragments would improve efficacy of PEP. Eight DNA samples were preamplified by PCR using different protocols. Treatments consisted of the use of Taq DNA polymerase (T), Taq plus a second polymerase obtained from Pyrococcus furiosus (E) or Stoffel fragment (S) in PEP. After preamplification, six genetic markers were genotyped, and the number of scorable genotypes was recorded. A control reaction (C) consisted of amplification using genomic DNA as template. A second experiment was performed to evaluate preamplification efficiency using Taq DNA polymerase (5 units) and exponential dilutions of Pfu DNA polymerase. After preamplification, the same procedure was used to obtain a number of scorable genotypes. In the first experiment, treatment E was the most reliable approach for amplifying genomic DNA in PEP. Treatments T and S produced fewer scorable genotypes than treatments E or C. In the second experiment, low concentrations of Pfu DNA polymerase produced a similar percentage of scorable genotypes as higher concentrations. Low concentrations of Pfu DNA polymerase combined with Taq DNA polymerase is the most cost-effective procedure to maximize amplification of limited DNA samples in PEP.

Animals

Development of bovine microsatellite markers from a microsatellite-enriched library.

A bovine genomic library enriched for DNA fragments bearing poly(dC-dA).poly(dG-dT) sequences was prepared and screened. Twenty-four clones bearing microsatellites were subject to sequence analysis and marker development as appropriate. Three of the 24 clones had microsatellites that were not evaluated for marker development owing to presence of a satellite element in two cases and limited repeat length in the third. Of the remaining 21 clones, all but one yielded polymorphic microsatellites. All but two of the polymorphic markers could be assigned to chromosomal locations by either linkage analysis or on the basis of X-linked inheritance as determined by heterozygosity limited to females. An unexpected clustering of markers was observed as 4 of the 20 polymorphic microsatellites mapped to a region of less than approximately 30 cM on Chromosome (Chr) 19, and four markers displayed X-linked inheritance.

Animals

A genetic linkage map of the bovine genome.

A cattle genetic linkage map was constructed which marks about 90% of the expected length of the cattle genome. Over 200 DNA polymorphisms were genotyped in cattle families which comprise 295 individuals in full sibling pedigrees. One hundred and seventy-one loci were found linked to one other locus. Twenty nine of the 30 chromosome pairs are represented by at least one of the 36 linkage groups. Less than a 50 cM difference was found in the male and female genetic maps. The conserved loci on this map show as many differences in gene order compared to humans as is found between humans and mice. The conservation is consistent with the patterns of karyotypic evolution found in the rodents, primates and artiodactyls. This map will be important for localizing quantitative trait loci and provides a basis for further mapping.

Animals