Annotation of the Affymetrix porcine genome microarray.
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Biomedical subjects
Publications and source records attributed to B A Freking.
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A resource population of ewes derived from Dorset and Romanov grandparents was generated over 5 yr to provide genotypic and phenotypic data to estimate effects of each callipyge genotype on component traits of reproduction. Effects on ovulation rate were estimated from data recorded by examination of ovaries from serially slaughtered ewe lambs (n = 174) born in 1994 and 1995. Productivity and longevity through 5 yr of age were recorded on a total of 265 F2 ewes produced in 1996 through 1998. Number of lambs born, birth weights, and weaning weights of lambs were calculated for each ewe. Weaning weight was recorded for each lamb and adjusted for age to 56 and 32 d for dam- and nursery-reared lambs, respectively. Longevity was determined as a binary trait based on the presence or absence of the ewe in the breeding flock at 5 yr of age. Ewes generated 937 records for breeding weight and 925 records for conception rate, which was determined on ewes exposed and still present at the start of each lambing season. Seven hundred seventeen records were analyzed for traits based on the number of ewes lambing. Genotypes for the causative callipyge mutation were determined using a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry assay. Each ewe was classified into 1 of 4 genotypes; NMatNPat, NMatCPat, CMatNPat, or CMatCPat, where N represents wild type and C represents the mutant allele. The mean ovulation rate of the ewe lambs genotyped NMatNPat was greater (P < 0.02) than that exhibited by NMatCPat; however, the estimated difference of 0.25 ovum in the ewe lambs did not affect the number born. The live weight of NMatCPat ewes at breeding was 2.5 kg less than that of other genotypes (P < 0.01). The callipyge genotype class did not significantly affect conception rates, fecundity, maternal ability, or ewe longevity. Maintaining a ewe flock to produce homozygous mutant rams for use in terminal sire mating systems presents no unusual problems from a reproduction standpoint.
There is concern about potential antagonistic correlated responses due to intensive selection for scrapie-resistant haplotypes of the prion (PRNP) gene in sheep. The objective of the present research was to test for associations of PRNP haplotypes for codons 136, 154, and 171 with growth, carcass, and meat quality traits in an F2 Dorset x Romanov population (n = 415) segregating the 2 callipyge alleles. Haplotypes of the 3 PRNP codons were determined for each sheep, and breed of origin of each gamete was predicted by genotyping 6 microsatellite markers flanking the PRNP locus. Twenty-five growth, carcass, and meat quality traits were evaluated. Data were analyzed using a basic model consisting of fixed effects of year, sex, and callipyge genotype, the random effect of sire, and 7 covariates corresponding to the probability that a lamb inherited a specific PRNP haplotype of either Dorset or Romanov origin. A fixed effect of litter size was added to the model for growth traits. The model for carcass traits contained the linear and quadratic effects of chilled carcass weight and the interactions among callipyge genotype and linear and quadratic terms. For meat quality traits, the model contained chilled carcass weight as a covariate and the interaction between callipyge genotype and chilled carcass weight. A contrast between the resistant ARR haplotype and the average effect of other PRNP haplotypes was tested to investigate the effects of potential selection for ARR within each breed of origin (Dorset, ARR vs. ARQ, VRQ, and AHQ; Romanov, ARR vs. ARQ and VRQ). There was limited evidence that selecting for scrapie resistance would cause correlated responses due to linkage disequilibrium. Associations of only 3 traits with PRNP haplotypes were detected in either breed of origin. In Romanov, the ARR haplotype was associated with longer carcasses (P < 0.013), narrower rumps (P = 0.038), and less marbling (P = 0.022) than the average of ARQ and VRQ haplotypes. No significant contrasts were detected for Dorset. This study is the first to account for breed of origin while investigating haplotype associations in an F2 population. This study provided limited evidence of associations between PRNP haplotypes and growth, carcass, and meat quality traits.
We hypothesized that the ability of the fetus to alter nutrient shunting and organ growth might be associated with uterine capacity. White crossbred gilts from a randomly selected control line, a line selected for ovulation rate, and a line selected for uterine capacity (UC) were unilaterally hysterectomized-ovariectomized at 160 d of age, mated at estrus, and slaughtered at 45, 65, 85, and 105 d of gestation (9 to 18 gilts for each line x day combination). Analysis of the data revealed that heart weights and fetal weights were decreased in the ovulation rate line. No significant differences were obtained in fetal, placental, or fetal organ weights between the control and UC lines. Allometric growth of organs was assessed by examination of the slopes of the relationships between fetal weights and fetal organ weights after natural log transformation. Only the relative growth of the liver differed between selection lines and was greater (P = 0.01) in the UC compared with the control line during early pregnancy (d 45 and 65). Allometric growth of the fetal brain, liver, and heart differed with day of gestation. A brain-sparing effect was greater (P < 0.01) on d 85 and 105 compared with d 45 and 65. By contrast, a heart-sparing effect was present during early gestation and disappeared in later gestation. Fetal liver weights were hypersensitive to differences in fetal weights on d 45, possibly associated with placental effects on fetal liver weight. Fetal spleen weights were proportional to fetal weights throughout gestation. These results indicate that selection for ovulation rate decreased total fetal and fetal heart weights, and that selection for UC altered the relationship between total fetal and fetal liver weights during early gestation. Results further indicate significant changes in allometric growth of organs during gestation.
A single nucleotide polymorphism (SNP; C vs. T) that creates an extra GATA-1 site (T allele) in intron 4 of the swine erythropoietin receptor (EPOR) gene was discovered and a genotyping assay for this SNP was developed. A total of 402 gilts from lines selected either at random (control), for ovulation rate (OR) or for uterine capacity (UC) for 11 generations were unilaterally hysterectomized-ovariectomized (UHO) at 160 days of age, mated at approximately 250 days of age and slaughtered at 105 days of pregnancy. Blood samples and spleens were collected from each foetus and the numbers of corpora lutea (CL) and live foetuses, the weights of each foetus and placenta, and each foetal haematocrit were recorded. In addition, intact gilts from the OR line or from a Yorkshire, Landrace, Duroc, crossbred line (BX) were mated and farrowed. At farrowing, the numbers of fully formed and live piglets were recorded for each litter. Genomic DNA was isolated for both the UHO and intact gilts, from foetuses from the UHO gilts that were heterozygous for the EPOR SNP, and from the boars from the BX line and were then used to determine EPOR SNP genotypes. Only CC and CT gilts were observed in the control, OR and UC selected lines. Presence of the EPOR T allele was associated (P < 0.05) with increased UC in these gilts. The number of heterozygous and homozygous foetuses did not differ within UHO litters, or did EPOR genotype influence foetal haematocrit. In intact gilts from the OR line, litter size was significantly associated (P < 0.05) with EPOR SNP genotype. Finally, results from intact gilts of the BX line, in which both the gilt and the boar genotypes were known, allowed an analysis to determine the effect of the gilt and/or the foetal genotype on litter size. This analysis indicated that the predicted foetal genotype (with gilt genotype as covariate) was associated with litter size (an increase of 2.6 +/- 1.0 piglets born alive predicted for homozygous T litters compared with homozygous C litters, P < 0.01) whereas the effect of the gilt genotype (adjusted for foetal genotype) on litter size was not significant. These results indicate that the EPOR SNP is associated with UC and litter size in two distinct populations and could be useful in increasing litter size in swine that are not limited in OR.
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Objectives were to estimate effects of sire breed (Dorset, Finnsheep, Romanov, Texel, and Montadale), dam breed [Composite III (CIII) and northwestern whiteface (WF)], mating season (March and May), and their interactions on reproductive traits of mature F1 ewes in spring mating seasons. A total of 1,099 F1 ewes produced 1,754 litters of 2,995 lambs from exposures to Suffolk rams during March and May mating seasons in 1995 through 1999. Fertility rate and ewe longevity were measured. Number born and litter birth weight were recorded, and number and weight at weaning and 20 wk of age were analyzed separately for dam- and nursery-reared litter mates. Total productivity from 4 to 6 yr of age for each ewe entering the breeding flock was calculated as the sum of 20-wk weights for dam-reared lambs and separately for nursery-reared lambs. Interactions of sire breed x mating season, ewe age x mating season, and ewe age x dam breed were often significant. Interactive effects of sire breed and mating season on fertility rate (P < 0.001) were primarily due to differences in magnitude. Fertility rates of sire breeds for March and May matings, respectively, were 92 and 89% for Romanov, 91 and 72% for Finnsheep, 90 and 52% for Texel, 88 and 52% for Montadale, and 83 and 62% for Dorset. Sire breed x mating season also affected number born (P < 0.03); March and May values were 2.12 and 2.05 for Romanov, 2.00 and 1.94 for Finnsheep, 1.39 and 1.41 for Texel, 1.37 and 1.51 for Montadale, and 1.37 and 1.55 for Dorset, respectively. Interaction of sire breed x dam breed on fertility rate (P < 0.01) was due to change in rank as well as magnitude. Romanov- and Dorset-sired ewes out of CIII dams had greater fertility rates than Romanov- and Dorset-sired ewes out of WF dams. The opposite situation existed for ewes by Finnsheep, Texel, and Montadale sires. Differences between dam breeds (CIII and WF) in total productivity of dam-reared lambs were not detected, whereas ewes exposed in March (78 kg) were more productive (P < 0.01) than those exposed in May (68 kg). Means of sire breeds for total productivity of dam-reared lambs were 47, 65, 70, 70, and 111 kg for Texel, Montadale, Dorset, Finnsheep, and Romanov, respectively (P < 0.001). Superior reproduction of Romanov sired ewes was primarily due to greater fertility rate and prolificacy at each mating season and ewe age. Use of Romanov-crossbred ewes would increase fertility during spring mating, an important constraint of the sheep industry.
Previous comparisons between the cDNA and gene sequences for secreted folate binding protein (sFBP) indicated a 12-bp insertion/deletion (ins/del) polymorphism in exon 1 and a SNP that altered (Ser-Arg) the protein AA sequence. The effect of the Ser-Arg SNP on reproductive traits was examined in three groups of Meishan-White European breed crossbred gilts. The gilts for all three groups were unilaterally hysterectomized-ovariectomized (UHO) at 100 d of age. Group 1 gilts (n = 77) were mated at estrus, slaughtered at d 105 of pregnancy, and a blood sample was collected from each fetus to determine fetal hematocrit. The number of corpora lutea and fetuses and the fetal and placental weights were recorded. Group 2 gilts (n = 46) were mated, the remaining uterine horn was flushed with 20 mL of saline on d 11 of pregnancy, conceptuses were counted, and flushings were measured for total sFBP. Gilts were allowed an estrous cycle to recover, mated again at estrus, slaughtered at 105 d of gestation, and the data as described for Group 1 were collected. Groups 1 and 2 gilts were genotyped for the Ser-Arg SNP. In Group 3, gilts (n = 70) and boars (n = 30) were genotyped for the Ser-Arg SNP before mating, and like genotypes were mated. Gilts were then treated as described for Group 2. The effect of the 12-bp ins/del on reproductive traits was examined in 407 white crossbred UHO gilts from a randomly selected control line and from lines selected for ovulation rate (OR) and uterine capacity (UC). Gilts were mated and slaughtered at 105 d of age, and the numbers of corpora lutea and live fetuses, and fetal and placental weights and fetal hematocrits were recorded. The 12-bp ins/del also was evaluated in 131 intact gilts from the OR selected line. These gilts were mated at approximately 250 d of age and farrowed. The numbers of fully formed and live piglets were recorded. A significant effect (P < 0.05) of the Ser-Arg SNP was detected on the number of embryos present on d 11 of pregnancy and on UC. The sFBP 12-bp ins/del was associated with UC (P < 0.01) and the number of CL (P < 0.05) in UHO gilts, but not with litter size in intact gilts from the OR line. Results suggest that the 12-bp ins/del polymorphism could be exploited to increase litter size in swine, provided that the negative effect of the polymorphism on OR is overcome.
The objectives of this study were to estimate effects of sire breed (Dorset, Finnsheep, Romanov, Texel, and Montadale), and dam breed (Composite III and Northwestern whiteface) on survival, growth, carcass, and composition traits of F1 lambs. Effects of mating season (August, October, and December) were estimated for survival and growth traits. Data were collected on 4,320 F1 lambs sired by 102 purebred rams over 3 yr. Birth weight was recorded on all lambs, and subsequent BW were adjusted to 56 (weaning), 70, and 140 d of age (n = 3,713, 3,654, and 3,579 observations, respectively). Survival of dam-reared progeny (n = 4,065) to weaning was recorded. Each year, wethers from October matings were slaughtered in three groups at 25, 29, and 33 wk of age to obtain carcass data (n = 546). In addition to standard carcass traits, resistive impedance measurements were recorded on the warm carcass to predict lean mass. Dam breed (P = 0.37) did not influence lamb survival to weaning, but sire breed (P < 0.05) was important. Romanov-sired lambs excelled in survival rate to weaning (94.1%), followed by Finn-sheep (93.0%), Texel (90.7%), Dorset (90.0%), and Montadale (89.1%) sired progeny. Lower (P < 0.01) postweaning growth rate was observed for Texel (267 g/d) and Finnsheep (272 g/d) sired progeny than for Dorset (285 g/d), Montadale (282 g/d), and Romanov (278 g/d) sired progeny. Sire breed and dam breed were generally significant for most carcass traits. Breed differences in distribution of carcass fat and carcass shape were detected; however, carcass composition was similar for all sire breeds when compared at a constant carcass weight. When evaluated at a constant 12th-rib fat depth, carcasses of lambs from Finnsheep, Romanov, and Texel sires produced 1 to 1.5 kg less (P < 0.001) predicted lean mass per lamb than carcasses of lambs from Dorset and Montadale sires. These experimental results provide information about the direct breed effects for survival, growth, and carcass traits of these breeds and their potential use in crossbreeding systems.
Objectives were to estimate effects of sire breed (Dorset, Finnsheep, Romanov, Texel, and Montadale), dam breed (Composite III and northwestern whiteface), mating season (August, October, and December), ewe age (1, 2, and 3 yr), and their interactions on reproductive traits of F1 ewes. A total of 1,799 F1 ewes produced 3,849 litters from 4,804 exposures to Suffolk rams during 35-d mating seasons over 3 yr. Ewes were weighed at breeding. Conception rate and ewe longevity (present or absent at 42 mo of age) were determined. Number born and litter birth weight were recorded, and number and weight at weaning and 20 wk of age were analyzed separately for dam- and nursery-reared litter mates. Total productivity through 3 yr of age for each ewe entering the breeding flock was calculated as the sum of 20-wk weights for dam- or nursery-reared lambs. Interactions of sire breed x mating season, sire breed x ewe age, and mating season x ewe age were generally significant, whereas interactions of sire breed, mating season, and ewe age x dam breed were seldom detected. Interactions of sire breed x mating season were often due to changes in rank as well as magnitude, indicating the importance of matching sire breed to a specific mating season. The number born to Dorset-, Texel-, and Montadale-sired ewes was not affected by dam breed; however, Finnsheep-sired ewes out of northwestern whiteface dams were more prolific than Finnsheep-sired ewes out of Composite III dams, and the opposite situation existed for Romanov-sired ewes. Least squares means of sire breeds (P < 0.001) for total productivity of dam-reared lambs were 98.5, 103.5, 106.9, 124.6, and 154.9 kg/ewe entering the breeding flock for Texel, Dorset, Montadale, Finnsheep, and Romanov, respectively. Superior reproduction of Romanov-sired ewes was due to greater conception rate and prolificacy for each mating season and ewe age, as well as greater ewe longevity. Total productivity of F1 ewes by Composite III dams (125.6 kg) was greater (P < 0.001) than for ewes born to northwestern whiteface dams (109.7 kg), and the effect of mating season increased (P < 0.001) from August to October to December. Litter weight at 20 wk of age of 2- and 3-yr-old ewes was similar but greater (P < 0.001) than for 1-yr-old ewes. Experimental results provide comprehensive information about the appropriate use of these breeds in crossbreeding systems to meet specific production-marketing objectives.
An experiment was designed to evaluate the effects of five sire breeds (Dorset, Finnsheep, Romanov, Texel, and Montadale), two dam breeds (Composite III [CIII] and northwestern whiteface [WF]), and three shearing seasons (December, February, and April, corresponding to August, October, and December breeding seasons) and their interactions on wool and other characteristics of F1 ewes. Fleeces were collected and characterized from six 2-yr-old F1 ewes representing each of the 90 sire breed x dam breed x shearing season x year (three) subclasses. Characteristics measured objectively were grease and clean fleece weights, clean yield, mean fiber diameter and SD, and mean staple length and SD. Visual assessments of fleece color were also made. Data collected on the F1 ewes were analyzed using a mixed model analysis of variance procedure. The model included fixed effects of year of birth, sire breed, dam breed, shearing season, six two-way interactions, and the three-way interaction of sire breed x dam breed x shearing season. The random effect of individual sire within year of birth x sire breed was also fitted. Texel- and Montadale-sired ewes produced more clean wool (P < 0.05) (approximately 0.24 kg) than Dorset-, Finnsheep-, and Romanov-sired ewes. Texel-sired ewes produced the coarsest wool (28.7 microm) (P < 0.05), whereas Romanov-sired ewes produced the finest (24.9 microm) and longest (9.12 cm) fleeces (P < 0.05). Ewes from WF dams produced more and finer wool (0.15 kg and 2.7 microm) than ewes from CIII dams (P < 0.001). Ewes shorn in December produced more, coarser, and longer wool (P < 0.05) than those shorn in February and April. This trend in wool production is opposite to that in conception rate (reported previously). Romanov-sired ewes produced the lowest percentage of white fleeces (62.6%), whereas Dorset-sired ewes produced the most (P < 0.001) white fleeces (96.3%). Estimates of heritability were calculated for grease and clean fleece weights (0.36), percentage of clean yield (0.31), average fiber diameter and SD (0.86 and 0.42, respectively), and average staple length and SD (0.49 and 0.00, respectively). Although necessary for a thorough evaluation of these 10 types of crossbred ewes, it is estimated that wool income would only constitute a small portion (1 to 5%) of overall income from sheep of this type.
High-throughput genotyping of swine populations is a potentially efficient method for establishing animal lineage and identification of loci important to animal health and efficient pork production. Markers were developed based upon single nucleotide polymorphisms (SNPs), which are abundant and amenable to automated genotyping platforms. The focus of this research was SNP discovery in expressed porcine genes providing markers to develop the porcine/human comparative map. Locus specific amplification (LSA) and comparative sequencing were used to generate PCR products and allelic information from parents of a swine reference family. Discovery of 1650 SNPs in 403 amplicons and strategies for optimizing LSA-based SNP discovery using alternative methods of PCR primer design, data analysis, and germplasm selection that are applicable to other populations and species are described. These data were the first large-scale assessment of frequency and distribution of porcine SNPs.
The objectives of this study were to assign both microsatellite and gene-based markers on porcine chromosome X to two radiation hybrid (RH) panels and to develop a more extensive integrated map of SSC-X. Thirty-five microsatellite and 20 gene-based markers were assigned to T43RH, and 16 previously unreported microsatellite and 15 gene-based markers were added to IMpRH map. Of these, 30 microsatellite and 12 gene-based markers were common to both RH maps. Twenty-two gene-based markers were submitted to BLASTN analysis for identification of orthologues of genes on HSA-X. Single nucleotide polymorphisms (SNPs) were detected for 12 gene-based markers, and nine of these were placed on the genetic map. A total of 92 known loci are present on at least one porcine chromosome X map. Thirty-seven loci are present on all three maps; 31 loci are found on only one map. Location of 33 gene-based markers on the comprehensive map translates into an integrated comparative map that supports conservation of gene order between SSC-X and HSA-X. This integrated map will be valuable for selection of candidate genes for porcine quantitative trait loci (QTLs) that map to SSC-X.
The starting point of the present study was the reported identification of a chromosomal region on bovine Chromosome (Chr) 15 (BTA15) carrying loci affecting meat tenderness. A comparative linkage map of BTA15 and human Chr 11 (HSA11) was constructed to identify potential positional candidate genes and to provide a resource of genetic markers to support marker-assisted selection (MAS). Relative rearrangements between the bovine and human genomes for these chromosomes are the most complex observed in comparative mapping between the two species, with nine alternating blocks of conserved synteny between HSA11 and bovine Chrs 15 and 29. The results of this study were the addition of nine genes to the HSA11/BTA15 comparative linkage map, and development of five microsatellite markers within the quantitative trait locus (QTL) interval. One gene with known effects on muscle development (MYOD1) was mapped to the interval. A second gene (CALCA) involved in regulation of calcium levels, a key factor in postmortem tenderization, also mapped within the interval. Refinement of the comparative map and QTL position will reduce the interval on the human transcription map to be scanned in search of candidates, reducing the effort and resources required to identify the allelic variation responsible for the genetic effect.
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An essential component of functional genomics studies is the sequence of DNA expressed in tissues of interest. To provide a resource of bovine-specific expressed sequence data and facilitate this powerful approach in cattle research, four normalized cDNA libraries were produced and arrayed for high-throughput sequencing. The libraries were made with RNA pooled from multiple tissues to increase efficiency of normalization and maximize the number of independent genes for which sequence data were obtained. Target tissues included those with highest likelihood to have impact on production parameters of animal health, growth, reproductive efficiency, and carcass merit. Success of normalization and inter- and intralibrary redundancy were assessed by collecting 6000-23,000 sequences from each of the libraries (68,520 total sequences deposited in GenBank). Sequence comparison and assembly of these sequences was performed in combination with 56,500 other bovine EST sequences present in the GenBank dbEST database to construct a cattle Gene Index (available from The Institute for Genomic Research at http://www.tigr.org/tdb/tgi.shtml). The 124,381 bovine ESTs present in GenBank at the time of the analysis form 16,740 assemblies that are listed and annotated on the Web site. Analysis of individual library sequence data indicates that the pooled-tissue approach was highly effective in preparing libraries for efficient deep sequencing.
A medium-density linkage map of the ovine genome has been developed. Marker data for 550 new loci were generated and merged with the previous sheep linkage map. The new map comprises 1093 markers representing 1062 unique loci (941 anonymous loci, 121 genes) and spans 3500 cM (sex-averaged) for the autosomes and 132 cM (female) on the X chromosome. There is an average spacing of 3.4 cM between autosomal loci and 8.3 cM between highly polymorphic [polymorphic information content (PIC) > or = 0.7] autosomal loci. The largest gap between markers is 32.5 cM, and the number of gaps of > 20 cM between loci, or regions where loci are missing from chromosome ends, has been reduced from 40 in the previous map to 6. Five hundred and seventy-three of the loci can be ordered on a framework map with odds of > 1000 : 1. The sheep linkage map contains strong links to both the cattle and goat maps. Five hundred and seventy-two of the loci positioned on the sheep linkage map have also been mapped by linkage analysis in cattle, and 209 of the loci mapped on the sheep linkage map have also been placed on the goat linkage map. Inspection of ruminant linkage maps indicates that the genomic coverage by the current sheep linkage map is comparable to that of the available cattle maps. The sheep map provides a valuable resource to the international sheep, cattle, and goat gene mapping community.
We used a comparative mapping approach to identify segments of conserved synteny between human Chromosome 14 (HSA14), bovine Chromosome 21 (BTA21), and the portion of ovine Chromosome 18 (OAR18) that contains the clpg locus. A bovine radiation hybrid map of the region was constructed with available Type II genetic markers and seven candidate genes to establish the comparative interval between BTA21 and HSA14. We developed polymorphic microsatellite and SNP markers associated with five candidate genes and placed them on the ovine and/or bovine genetic maps by multipoint linkage analysis. Three additional genes were mapped by virtue of their physical linkage to genetically mapped makers. Development of integrated linkage and physical maps facilitates the selection of positional candidate genes from the gene rich human map. The physically linked candidate genes PREF-1 and MEG3 map to the interval containing the clpg locus. Comparative biology suggests imprinting of MEG3 and/or the influences of PREF-1 on cellular differentiation, should be examined for their role in the parent-of-origin dependent influence of mutant clpg alleles on sheep muscle characteristics.