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B W Kirkpatrick

Publications and source records attributed to B W Kirkpatrick.

At least 55 records · Page 3Linked to original sources

Conformation polymorphisms and targeted marker development.

Genetic markers were developed for three targeted bovine loci using both single-stranded and double-stranded DNA conformation polymorphisms. Eight of nine DNA fragments exhibited single-stranded conformation polymorphisms while only one of nine exhibited a double-strand conformation polymorphism. All but one of the polymorphic fragments exhibited two allelic forms, with the exception being a single-stranded conformation polymorphism with three alleles. Utility of conformation polymorphisms relative to microsatellite markers for linkage map development and quantitative trait loci (QTL) mapping was assessed by comparing frequency of heterozygote parents from reference and resource families. Heterozygosity was greater for microsatellite markers (P < 0.01) and reference family parents (P < 0.01), though the disparity between marker types tended to be less dramatic for the reference family parents (P < 0.09). These results suggest conformation polymorphisms will be a useful tool for targeted marker development in linkage map and comparative map development provided genetically diverse families are studied.

Animals↗

A genetic map of index DNA loci on bovine chromosome 21.

Genetic maps of highly polymorphic index DNA loci exist essentially only in humans and some experimental organisms. Here we report the first genetic map of highly polymorphic index DNA loci in livestock for bovine chromosome 21. This map consists of six loci with an average heterozygosity of 82%, each with a minimum of five alleles, spaced at an average genetic distance of 9.7 cM, and covers most of the expected length of the acrocentric bovine chromosome 21. The order of markers along the chromosome is cen-ETH 131-UWCA 4-TGLA 337-TGLA 122-CSSM 18-GMBT 16-tel. There is heterogeneity among the recombination fractions between the sexes.

Animals↗

Double-strand DNA conformation polymorphisms as a source of highly polymorphic genetic markers.

The molecular basis for several apparent restriction fragment length polymorphisms of the porcine growth hormone gene was examined through DNA sequence analysis. Electrophoretic and sequence analysis suggest polymorphisms result from 1-3 base substitutions that affect double-strand DNA conformation and electrophoretic mobility. Two allelic forms of the porcine growth hormone 5' flank and four allelic forms of the second exon/intron region were identified. A marker system was developed which combined conformation polymorphisms with HaeII and DdeI RFLPs. Using this system, nine haplotypes were observed in samples from three US swine breeds. The data presented suggest that double-strand DNA conformation can be exploited in base substitution detection and development of highly polymorphic genetic marker systems.

Alleles↗

Sensitive sex determination assay applicable to bovine embryos derived from IVM and IVF.

A rapid, sensitive method for sex determination which uses nested, allele-specific amplification of bovine zfx and zfy was developed. Accuracy of the sex determination assay was verified with lymphocyte-derived DNA from 24 cattle of known gender. Reliability and sensitivity of the assay in embryonic sex determination were examined by typing duplicate bovine embryo biopsies of 2-8 cells. Sex determination was possible in 38 of 40 biopsies, and agreement between biopsy assays occurred in 17 of 18 cases. Applicability of the sex determination assay was further verified by examining gender of 12 fetuses derived from sexed embryos. Embryonic and fetal gender determinations agreed in 11 of 12 cases with the twelfth yielding an ambiguous result from fetal ultrasound examination.

Animals↗

Olfactory bulb removal affects partner preference development and estrus induction in female prairie voles.

In female prairie voles (Microtus ochrogaster) bilateral olfactory bulbectomy reduced affiliative behavior, as measured by social contact, and prevented the formation of partner preferences. Unilateral olfactory bulb removal did not significantly influence affiliative behavior, but did inhibit partner preferences. Bilateral, but not unilateral, bulbectomy significantly reduced the proportion of females exhibiting behavioral estrus following male exposure. In contrast to affiliative and sexual behavior, parental behavior was not significantly affected by either bilateral or unilateral olfactory bulbectomy. These results suggest that divergent sensory-neural pathways underlie social, sexual, and parental behaviors in this species.

Aggression↗

Identification of a conserved microsatellite site in the porcine and bovine insulin-like growth factor-I gene 5' flank.

Examination of published rat and human sequences for the insulin-like growth factor-I (IGF-I) gene indicated the presence of CA dinucleotide repeats in corresponding segments of each. Presence of similar microsatellite sequences in the porcine and bovine IGF-I genes was hypothesized. A 1200-bp segment upstream of the porcine and bovine IGF-I genes was amplified using the polymerase chain reaction (PCR) with primers developed from a consensus of human, rat and bovine sequences. Both porcine and bovine PCR products contained similar microsatellite sequences. Amplified pIGF-I DNA was cloned and sequenced, and an additional primer was developed specifically for microsatellite marker detection. Six allelic variants of 124, 130, 132, 134, 136 or 138 bp were observed in pigs with differing frequencies between breeds (P < 0.01). The same primers were used to amplify the corresponding bovine microsatellite. Three alleles of 126, 128 and 130 bp were observed in a genetically diverse cattle population with estimated frequencies of 0.06, 0.68 and 0.26, respectively. Results of this study indicate sequence information from the human and laboratory species can be used to facilitate genetic marker development in livestock species.

Alleles↗

The effect of photoperiod on diurnal rhythms of serum gonadotrophins, prolactin and melatonin in ovariectomized heifers.

Fourteen heifers, ovariectomized prepubertally, were used in two experiments to test the hypotheses that (1) there are diurnal patterns in circulating concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin and melatonin and (2) that these diurnal patterns would be altered by changing photoperiod. In experiment 1 (Exp. 1) animals were randomly assigned to either increasing photoperiod (I) (n = 7) or decreasing photoperiod (D) (n = 7) for a period of 8 weeks. In the second experiment (Exp. 2) four heifers from each photoperiod group were assigned to the opposite treatment for another 4 weeks forming four subgroups; continued increasing photoperiod (II) (n = 3), increasing followed by decreasing photoperiod (ID) (n = 4), decreasing followed by increasing photoperiod (DI) (n = 4) and continued decreasing photoperiod (DD) (n = 3). At weeks 4, 8 (Exp. 1) and 12 (Exp. 2) the animals were cannulated and blood samples were taken hourly from 1030 hr to 0830 hr (23 samples). In Exp. 1, animals exposed to either I or D did not differ in circulating concentrations of LH, FSH, prolactin and melatonin. Concentrations of LH and melatonin but not FSH or prolactin were higher in animals during the scotophase than the photophase. In Exp. 2 LH, FSH and prolactin, but not melatonin concentrations were higher in animals exposed to I (II and ID) than D (DI and DD). Animals exposed to I had higher circulating concentrations of FSH and animals exposed to both I and D had higher concentrations of melatonin during the scotophase. Neither LH nor prolactin concentrations differed between scotophase and photophase. The abrupt switch from one photoperiod treatment to the other did not significantly affect LH, FSH or prolactin but did alter the patterns and concentrations of circulating melatonin. Animals that remained in the same photoperiod treatment had similar melatonin concentrations; but animals changed from I to D had higher serum melatonin concentrations than those changed from D to I. These data support the hypothesis that there are diurnal changes in circulating concentrations of melatonin in the prepubertal bovine female, with concentrations higher during scotophase than photophase. In addition, under certain photoperiodic conditions there were diurnal rhythms in gonadotropins. These data also support the hypothesis that changing photoperiod alters the diurnal pattern in circulating melatonin concentrations.

Animals↗