Quantitative trait locus analysis of plasma cholesterol levels and body weight by controlling the effects of the Apoa2 allele in mice

Colleagues and I previously performed quantitative trait locus (QTL) analysis on plasma total-cholesterol (T-CHO) levels in C57BL/6J (B6) x RR F2 mice. We identified only one significant QTL (Cq6) on chromosome 1 in a region containing the Apoa2 gene locus, a convincing candidate gene for Cq6. Because Cq6 was a highly significant QTL, we considered that the detection of other potential QTLs might be hindered. In the present study, QTL analysis was performed in B6.KK-Apoa2b N(8) x RR F2 mice [B6.KK-Apoa2b N(8) is a partial congenic strain carrying the Apoa2b allele from the KK strain, and RR also has the Apoa2b allele] by controlling of the effects of the Apoa2 allele, for identifying additional QTLs. Although no significant QTLs were identified, 2 suggestive QTLs were found on chromosomes 2 and 3 in place of the effects of the Apoa2 allele. A significant body weight QTL was identified on chromosome 3 (Bwq7, peak LOD score 5.2); its effect on body weight was not significant in previously analyzed B6 x RR F2 mice. Suggestive body weight QTL that had been identified in B6 x RR F2 mice on chromosome 4 (LOD score 3.8) was not identified in B6.KK-Apoa2b N(8) x RR F2 mice. Thus, contrary to expectation, the genetic control of body weight was also altered significantly by controlling of the effects of the Apoa2 allele. The QTL mapping strategy by controlling of the effects of a major QTL facilitated the identification of additional QTLs.     

Characterization of Cq3, a quantitative trait locus that controls plasma cholesterol and phospholipid levels in mice

Cq3 was identified in C57BL/6J (B6) x KK-Ay F2 mice as a quantitative trait locus (QTL) that controls plasma cholesterol and phospholipid levels, and normolipidemic B6 allele was associated with increased lipids. Cq3 was statistically significant in F2-a/a, but not in F2-Ay/a; probably because the Cq3 effect was obscured by introduction of the Ay allele, which in itself has a strong hyperlipidemic effect. Because the peak LOD score for Cq3 was identified near D3Mit102 (49.7 cM) on chromosome 3, linkage analyses with microsatellite markers located at 49.7 cM were performed in KK x RR F2, B6 x RR F2, and KK x CF1 F2. However, even a suggestive QTL was not identified in any of the three F2. By testing all pairs of marker loci, I found a significant interaction between Cq3 and the Apoa2 locus, and F2 mice with the Apoa2(KK)/Apoa2(KK); D3Mit102(B6)/D3Mit102(B6) genotype had significantly higher cholesterol levels than did F2 mice with other genotypes. The results showed that the ;round-robin' strategy was not always applicable to the search for QTL genes; probably because specific gene-to-gene interaction limited the validity of the strategy to the utmost extent.     

Quantitative trait loci that control plasma lipid levels in an F2 intercross between C57BL/6J and DDD.Cg-A(y) inbred mouse strains

The objectives of this study were to characterize plasma lipid phenotypes and dissect the genetic basis of plasma lipid levels in an obese DDD.Cg-A(y) mouse strain. Plasma triglyceride (TG) levels were significantly higher in the DDD.Cg-A(y) strain than in the B6.Cg-A(y) strain. In contrast, plasma total-cholesterol (CHO) levels did not substantially differ between the two strains. As a rule, the A(y) allele significantly increased TG levels, but did not increase CHO levels. Quantitative trait locus (QTL) analyses for plasma TG and CHO levels were performed in two types of F(2) female mice [F(2)A(y) (F(2) mice carrying the A(y) allele) and F(2) non- A(y) mice (F(2) mice without the A(y) allele)] produced by crossing C57BL/6J females and DDD.Cg-A(y) males. Single QTL scan identified one significant QTL for TG levels on chromosome 1, and two significant QTLs for CHO levels on chromosomes 1 and 8. When the marker nearest to the QTL on chromosome 1 was used as covariates, four additional significant QTLs for CHO levels were identified on chromosomes 5, 6, and 17 (two loci). In contrast, consideration of the agouti locus genotype as covariates did not detect additional QTLs. DDD.Cg-A(y) showed a low CHO level, although it had Apoa2(b), which was a CHO-increasing allele at the Apoa2 locus. This may have been partly due to the presence of multiple QTLs, which were associated with decreased CHO levels, on chromosome 8.     

Molecular characterization of avian polyomavirus isolated from psittacine birds based on the whole genome sequence analysis

Seven avian polyomaviruses (APVs) were isolated from seven psittacine birds of four species. Their whole genome sequences were genetically analyzed. Comparing with the sequence of BFDV1 strain, nucleotide substitutions in the sequences of seven APV isolates were found at 63 loci and a high level of conservation of amino acid sequence in each viral protein (VP1, VP2, VP3, VP4, and t/T antigen) was predicted. An A-to-T nucleotide substitution was observed in non-control region of all seven APV sequences in comparison with BFDV1 strain. Two C-to-T nucleotide substitutions were also detected in non-coding regions of one isolate. A phylogenetic analysis of the whole genome sequences indicated that the sequences from the same species of bird were closely related. APV has been reported to have distinct tropism for cell cultures of various avian species. The present study indicated that a single amino acid substitution at position 221 in VP2 was essential for propagating in chicken embryonic fibroblast culture and this substitution was promoted by propagation on budgerigar embryonic fibroblast culture. For two isolates, three serial amino acids appeared to be deleted in VP4. However, this deletion had little effect on virus propagation.     

Detection of quantitative trait loci for fat androstenone levels in pigs

A QTL analysis of fat androstenone levels from a three-generation experimental cross between Large White and Meishan pig breeds was carried out. A total of 485 F2 males grouped in 24 full-sib families, their 29 parents and 12 grandparents were typed for 137 markers distributed over the entire porcine genome. The F2 male population was measured for fat androstenone levels at 100, 120, 140, and 160 d of age and at slaughter around 80 kg liveweight. Statistical analyses were performed using two interval mapping methods: a line-cross (LC) regression method, which assumes alternative alleles are fixed in founder lines, and a half- full-sib (HFS) maximum likelihood method, where allele substitution effects were estimated within each half- and full-sib family. Both methods revealed genomewide significant gene effects on chromosomes 3, 7, and 14. The QTL explained, respectively, 7 to 11%, 11 to 15%, and 6 to 8% of phenotypic variance. Three additional significant QTL explaining 4 to 7% of variance were detected on chromosomes 4 and 9 using LC method and on chromosome 6 using HFS method. Suggestive QTL were also obtained on chromosomes 2, 10, 11, 13, and 18. Meishan alleles were associated with higher androstenone levels, except on chromosomes 7, 10, and 13, although 10 and 13 additive effects were near zero. The QTL had essentially additive effects, except on chromosomes 4, 10, and 13. No evidence of linked QTL or imprinting effects on androstenone concentration could be found across the entire porcine genome. The steroid chromosome P450 21-hydroxylase (CYP21) and cytochrome P450 cholesterol side chain cleavage subfamily XIA (CYP11A) loci were investigated as possible candidate genes for the chromosome 7 QTL. No mutation of coding sequence has been found for CYP21. Involvement of a candidate regulatory mutation of CYP11A gene proposed by others can be excluded in our animals.     

Evaluation of single-nucleotide polymorphisms in CAPN1 for association with meat tenderness in cattle

Micromolar calcium activated neutral protease (CAPN1) was evaluated as a candidate gene for a quantitative trait locus (QTL) on BTA29 affecting meat tenderness by characterization of nucleotide sequence variation in the gene. Single-nucleotide polymorphisms (SNP) were identified by sequencing all 22 exons and 19 of the 21 introns in two sires (Piedmontese x Angus located at the U.S. Meat Animal Research Center in Clay Center, NE; Jersey x Limousin located at AgResearch in New Zealand) of independent resource populations previously shown to be segregating meat tenderness QTL on BTA29. The majority of the 38 SNP were found in introns or were synonymous substitutions in the coding regions, with two exceptions. Exons 14 and 9 contained SNP that were predicted to alter the protein sequence by the substitution of isoleucine for valine in Domain III of the protein, and alanine for glycine in Domain II of the protein. The resource populations were genotyped for these two SNP in addition to six intronic polymorphisms and two silent substitutions. Analysis of genotypes and shear force values in both populations revealed a difference between paternal CAPN1 alleles in which the allele encoding isoleucine at position 530 and glycine at position 316 associated with decreased meat tenderness (increased shear force values) relative to the allele encoding valine at position 530 and alanine at position 316 (P < 0.05). The association of maternal alleles with meat tenderness phenotypes is consistent with the hypothesis of CAPN1 as the gene underlying the QTL effect in two independent resource populations and presents the possibility of using these markers for selective breeding to reduce the numbers of animals with unfavorable meat tenderness traits.     

Genome-wide linkage and QTL mapping in porcine F2 families generated from Pietrain, Meishan and Wild Boar crosses

Three informative pig F₂ families based on European Wild Boar (W), Meishan (M) and Pietrain (P) crosses have been used for genome‐wide linkage and quantitative trait loci (QTL) analysis. Altogether 129 microsatellites, 56 type I loci and 46 trait definitions (specific to growth, fattening, fat deposition, muscling, meat quality, stress resistance and body conformation) were included in the study. In the linkage maps of M × P, W × P and W × M families, average spacing of markers were 18.4, 19.7 and 18.8 cM, the numbers of informative meioses were 582, 534 and 625, and the total lengths of autosomes measured were 27.3, 26.0 and 26.2 Morgan units, respectively. Maternal maps were on average 1.3 times longer than paternal maps. QTLs contributing more than 3% of F₂ phenotypic variance could be identified at p < 0.05 chromosome‐wide level. Differences in the numbers and positions of QTLs were observed between families. Genome‐wide significant QTL effects were mapped for growth and fattening traits on eight chromosomes (1, 2, 4, 13, 14, 17, 18 and X), for fat deposition traits on seven chromosomes (1, 2, 3, 4, 6, 7 and X), for muscling traits on 11 chromosomes (1, 2, 3, 4, 6, 7, 8, 12, 14, 15 and X), for meat quality and stress resistance traits on seven chromosomes (2, 3, 6, 13, 16, 18 and X), and QTLs for body‐conformation traits were detected on 14 chromosomes. Closely correlated traits showed similar QTL profiles within families. Major QTL effects for meat quality and stress resistance traits were found on SSC6 in the interval RYR1‐A1BG in the W × P and M × P families, and could be attributed to segregation of the RYR1 allele T derived from Pietrain, whereas no effect in the corresponding SSC6 interval was found in family W × M, where Wild Boar and Meishan both contributed the RYR1 allele C. QTL positions were mostly similar in two of the three families for body conformation traits and for growth, fattening, fat deposition and muscling traits, especially on SSC4 (interval SW1073‐NGFB). QTLs with large effects were also mapped on SSC7 in the major histocompatibility complex (MHC) (interval CYP21A2‐S0102) and affected body length, weight of head and many other traits. The identification of DNA variants in genes causative for the QTLs requires further fine mapping of QTL intervals and a positional cloning. However, for these subsequent steps, the genome‐wide QTL mapping in F₂ families represents an essential starting point and is therefore significant for animal breeding.     

玉米持绿相关ＱＴＬ整合图谱构建及一致性ＱＴＬ区域内候选基因发掘

 以玉米高密度遗传连锁图谱ＩＢＭ２２００８Ｎｅｉｇｈｂｏｒｓ为参考图谱,收集来自不同实验中的１７３个玉米持绿相关数量性状位点（ｑｕａｎｔｉｔａｔｉｖｅｔｒａｉｔｌｏｃｕｓ,ＱＴＬ）信息,利用ＢｉｏＭｅｒｃａｔｏｒ２．１软件,构建出玉米持绿相关ＱＴＬ 整合图谱;采用元分析技术,在１,４,５,９号染色体上发掘出５个持绿“一致性”ＱＴＬ 区间。根据“一致性”ＱＴＬ 区间两端标记在玉米物理图谱Ｂ７３ＲｅｆＧｅｎ＿ｖ２上的位置,将“一致性”ＱＴＬ 区间进行物理图谱定位,利用ＰｌａｎｔＧＤＢ（ｈｔｔｐ：／／ｗｗｗ．ｐｌａｎｔｇｄｂ．ｏｒｇ／）在线区段批量下载工具（ｄｏｗｎｌｏａｄｒｅｇｉｏｎｄａｔａ）下载“一致性”区间的１４４５个预测基因序列并进行生物信息学分析,发现预测基因主要参与具体的细胞过程,执行结合功能,催化、转移酶活性和氧化还原酶活性等分子功能。根据“一致性”ＱＴＬ 区间的基因位点名称,在ＮＣＢＩ中下载相关基因序列,与所在“一致性”ＱＴＬ 区 间所有预测基因保守结构域进行比对,在５个“一致性”持绿ＱＴＬ 区间内初步确定８个持绿相关候选基因。利用ＧＲＡＭＥＮＥ 网站（ｈｔｔｐ：／／ｗｗｗ．ｇｒａｍｅｎｅ．ｏｒｇ／）的Ｃｍａｐ功能,将水稻持绿基因狊犵狉（ｓｔａｙｇｒｅｅｎ）转定位于玉米物理图谱Ｂ７３ＲｅｆＧｅｎ＿ｖ２上,找到与其同源的玉米候选基因ＧＲＭＺＭ２Ｇ０９１８３７＿Ｔ０１,其序列与已发表的玉米衰老诱导叶绿体持绿蛋白基因狊犵狉１序列一致。     

Single- and joint-population analyses of two experimental pig crosses to confirm quantitative trait loci on Sus scrofa chromosome 6 and leptin receptor effects on fatness and growth traits

The primary goal of this study was to detect and confirm QTL on SSC6 for growth and fatness traits in 2 experimental F(2) intercrosses: Iberian x Landrace (IB x LR) and Iberian x Meishan (IB x MS), which were used in this study for the first time in a QTL analysis related to productive traits. For this purpose, single- and joint-population analyses with single and bivariate trait models of both populations were performed. The presence of the SSC6 QTL for backfat thickness previously identified in the IB x LR cross was detected in this population with additional molecular information, but also was confirmed in the IB x MS cross. In addition, a QTL affecting BW was detected in both crosses in a similar position to the QTL detected for backfat thickness. This is the first study in which a QTL affecting BW is detected on SSC6 in the IB x LR cross, as well as in the IB x MS resource population. Furthermore, we analyzed a previously described nonsynonymous leptin receptor (LEPR) SNP located in exon 14 (c.2002C > T) for causality with respect to this QTL within both F(2) populations. Our results supported the previously reported association between LEPR alleles and backfat thickness in the IB x LR cross, and this association was also confirmed within the IB x MS cross. An association not reported before between LEPR alleles and BW was identified in both populations.     

紫花苜蓿粗灰分与矿质元素含量QTL定位分析

为初步鉴定与紫花苜蓿(Medicago sativa)粗灰分、钾、钙、镁、磷含量调控相关的数量性状基因座(Quantitative trait loci, QTL)和分子标记,本研究用低产早熟苜蓿和高产晚熟苜蓿杂交,构建了由392个个体组成的F₁群体,对这些性状进行3年表型数据测定,且基于前期已构建的高密度遗传连锁图谱开展QTL定位。结果表明：共检测到63个与粗灰分和4种矿质元素含量相关的QTL,分布于22个染色体上,单个QTL的贡献率为2.50%～29.85%;其中重复定位的QTL共8个(qCa-3C-1和qCa-3C-2,qCa-6B-1和qCa-6B-2,qCa-6D-1和qCa-6D-2,qAsh-8B-1和qAsh-8B-2),共定位的QTL有6个(qP-2C和qK-2C,qP-3A和qMg-3A,qP-6D-3和qMg-6D-2);经进一步验证,与这些QTL紧密连锁的标记可用于分子标记辅助选择育种。本研究为选育矿质营养更丰富的苜蓿新品种奠定了基础。     

A quantitative trait locus genome scan for porcine muscle fiber traits reveals overdominance and epistasis

Muscle histochemical characteristics are decisive determinants of meat quality. The relative percentage and diameters of the different muscular fiber types influence crucial aspects of meat such as color, tenderness, and ultimate pH. Despite its relevance, however, the information on muscle fiber genetic architecture is scant, because histochemical muscle characterization is a laborious task. Here we report a complete QTL scan of muscle fiber traits in 160 animals from a F(2) cross between Iberian and Landrace pigs using 139 markers. We identified 20 genome regions distributed along 15 porcine chromosomes (SSC1, 2, 3, 4, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, and X) with direct and(or) epistatic effects. Epistasis was frequent and some interactions were highly significant. Chromosomes 10 and 11 seemed to behave as hubs; they harbored 2 individual QTL, but also 6 epistatic regions. Numerous individual QTL effects had cryptic alleles, with opposite effects to phenotypic pure breed differences. Many of the QTL identified here coincided with previous reports for these traits in the literature, and there was overlapping with potential candidate genes and previously reported meat quality QTL.     

Genetic analysis of litter size in mice

We performed quantitative trait locus (QTL) mapping analysis for litter size (total number of pups born and/or number of pups born alive) in 255 backcross mice derived from C57BL/6J and RR/Sgn inbred mice. We identified one significant QTL on chromosome 7 and 4 suggestive QTLs on chromosomes 3, 5, 10 and 13. In addition, two suggestive QTLs were identified on chromosomes 1 and 4 for the number of stillbirth. These results suggested that both litter size and number of stillbirth were heritable traits, although they were controlled by distinct genes. The RR allele was associated with reduced litter size and increased stillbirth at all QTLs. Therefore, RR mothers were observed to have reduced prolificacy in this particular genetic cross.     

水稻饲料营养含量的QTL定位分析

应用85个SSR标记,对普通野生稻与粳稻台中65为亲本建立的F₂群体进行基因检测,构建了覆盖水稻基因组12条染色体的SSR分子标记连锁图,采用Mapmaker/QTL1.0统计软件对决定水稻饲用营养价值的粗蛋白、粗纤维、粗脂肪、粗灰分、硅酸和可溶性糖含量的基因座位进行了定位分析。结果定位了影响粗蛋白含量的3个QTLs,影响粗脂肪含量的1个QTL,影响可溶性糖含量的3个QTLs,影响硅酸含量的2个QTLs,这9个QTLs分别位于第1,2,4,7,8,9,10和11染色体上。其中主效QTL4个,分别是影响粗脂肪含量的qCEE-1(贡献率56.8%),影响可溶性糖含量的qCWSC-4(贡献率23.1%)和qCWSC-7(贡献率25.0%),影响硅酸含量的qCS-9(贡献率15.9%),其余5个为微效QTL。没有检测到影响粗纤维含量和粗灰分含量的QTL。     

Genetic variation of major histocompatibility complex BLB2 gene exon 2 in Hebei domestic chicken

Genetic variation of MHC BLB2 gene exon 2 in Hebei domestic chicken was investigated, after PCR and sequencing of a 374bp fragment (containing entire exon 2 (270bp) of BLB2 gene) in 76 individuals. The results showed that along this fragment, there were 69 variable sites, of which 18 were novel variations, and 82 estimated haplotypes with the diversity of 0.960. In Hebei domestic chicken, the nucleotide diversity (π), the average number of nucleotide differences (k), the average number of nucleotide diversity of synonymous substitution (π(s)) and non-synonymous substitution (π(a)) in BLB2 gene exon 2 were 0.098, 24.688, 0.075, and 0.106, respectively; nine non-synonymous substitutions was exclusively found in the peptide-binding sites (PBS) region of BLB2 gene exon 2, inferring that these unique substitutions might be helpful to resist some special bacteria and pathogens. The higher genetic diversity of MHC BLB2 gene exon 2 in Hebei domestic chicken might be consistent with its more robust disease resistance.     

Mitochondrial genome polymorphisms associated with longissimus muscle composition in Iberian pigs

We carried out a study to investigate the associations between mitochondrial DNA polymorphisms and meat quality traits (intramuscular fat and protein content of the longissimus) in an Iberian porcine line named Torbiscal. The studied pigs (n = 319) belong to 9 maternal lineages and were previously assigned to 6 mitochondrial haplotypes (H1 to H6), based on Cytochrome b and Dloop sequences. Statistical analyses, following a bivariate mixed model, show a greater fat content and lower protein content in H3 haplotype carriers than H1, H2, H4, H5, and H6 haplotype carriers. The magnitudes of these differences are close to 1 g of fat and -0.5 g of protein per 100 g of muscle. To identify the causative mutation of these effects on intramuscular fat and protein contents, the complete mitochondrial DNA sequence of 6 individuals was determined, each one carrying a different mitochondrial haplotype. The alignments of these 6 complete mitochondrial sequences allowed identification of 32 substitutions and 2 indels. Two polymorphic positions were exclusively detected in H3 carriers: a synonymous transition 9104C > T in the gene-coding region of Cytochrome c oxidase subunit III and a substitution 715A > G in 12S rRNA. Genotyping results of a larger number of Torbiscal samples showed the exclusive presence of 9104T and 715G alleles in H3 carriers. The detected candidate substitutions are located in essential mitochondrial genes, and although they do not change the amino acid composition, we cannot disregard a potential change in the secondary structure of their corresponding mRNA. The usefulness of these polymorphisms as markers in selection programs requires validation of the consistency of these results in other Iberian pig lines.     

Detection and fine mapping of quantitative trait loci for bone traits on chicken chromosome one

In broiler chickens, bone problems are an important welfare issue that has been linked to genetic selection for rapid growth. The objectives of this study were to identify and fine map quantitative trait loci (QTL) associated with bone traits. The Northeast Agricultural University resource population (NEAURP) being an F(2) population was used in this study, and a total of 17 bone traits were measured. In primary genome scan, the linkage map was constructed with 23 microsatellite markers across the entire chicken chromosome 1. Seventeen QTLs for bone traits were identified and 12 of these were found between LEI0079 and ROS0025 (50.8 cM apart). To fine map the QTLs located between LEI0079 and ROS0025, more markers and more individuals were used and a new partial linkage map was constructed. The confidence intervals for QTLs were sharply narrowed down from 24.5～52.6 to 2.7～17.0 Mb. This study identified chromosome regions harbouring significant QTLs affecting bone traits and showed that the use of more markers and individuals could decrease the confidence interval of QTL effectively. The results provide a useful reference for further candidate gene research and MAS for bone traits.     

Fine mapping of quantitative trait loci affecting organ weights by mouse intersubspecific subcongenic strain analysis

The genetic architecture of organ weights is not well understood. In this study, we fine‐mapped quantitative trait loci (QTLs) affecting organ weights by characterizing six intersubspecific subcongenic mouse strains with overlapping and non‐overlapping genomic regions on chromosome 2 derived from wild Mus musculus castaneus. QTLs for heart, lung, spleen and kidney weights were revealed on a 6.38‐Mb genomic region between two microsatellite markers, D2Mit323 and D2Mit472. Effects of the castaneus alleles at the organ weight QTLs were all opposite in direction to a body weight QTL previously mapped to the same genomic region. In addition, new QTLs for lung and kidney weights were revealed on a different 3.57‐Mb region between D2Mit205 and D2Mit182. Their effects were dependent on that of another body weight QTL previously mapped to that genomic region. The organ weight QTLs revealed were all duplicated in independent analyses with F₂ intercross populations between subcongenic strains carrying these QTLs and their background strain. The findings suggested that organ weights are not exclusively regulated by genetic loci that commonly influence overall body weight and rather that there are loci contributing to the growth of specific organs only.     

Genetic mapping of the QTL affecting body weight in chickens using a F2 family

1. To identify the quantitative trait loci (QTL) affecting growth in chickens, we carried out QTL analysis on chicken growth traits using a population of 227 F2 crosses between a Satsumadori (slow-growing, light-weight Japanese native breed used as a meat chicken) male and a White Plymouth Rock (early-maturing, heavy weight broiler). 2. We chose 78 microsatellite loci from 331 publicly available on 14 linkage groups, with respect to their utility and location. 3. Two QTLs affecting body weight at 13 and 16 weeks were mapped at 220 cM on chromosome 1 (LOD scores, 2.8 and 4.5, respectively, at 13 and 16 weeks), and at 60 cM on chromosome 2 (LOD scores, 6.2 and 8.1, respectively, at 13 and 16 weeks). 4. The closest loci to the QTLs were LEI71 on chromosome 1 and LMU13 and MCW184 on chromosome 2. 5. The sites of the QTLs agreed closely with those already reported. Therefore, it seems likely that QTLs affecting growth of chickens are located at these sites.     

Fine mapping a quantitative trait locus affecting ovulation rate in swine on chromosome 8

Ovulation rate is an integral component of litter size in swine, but is difficult to directly select for in commercial swine production. Because a QTL has been detected for ovulation rate at the terminal end of chromosome 8p, genetic markers for this QTL would enable direct selection for ovulation rate in both males and females. Eleven genes from human chromosome 4p16-p15, as well as one physiological candidate gene, were genetically mapped in the pig. Large insert swine genomic libraries were screened, clones were isolated and then screened for microsatellite repeats, and informative microsatellite markers were developed for seven genes (GNRHR, IDUA, MAN2B2, MSX1, PDE6B, PPP2R2C, and RGS12). Three genes (LRPAP1, GPRK2L, and FLJ20425) were mapped using genotyping assays developed from single nucleotide polymorphisms. Two genes were assigned since they were present in clones that contained mapped markers (HGFAC and HMX1). The resulting linkage map of pig chromosome 8 contains markers associated with 14 genes in the first 27 cM. One inversion spanning at least 3 Mb in the human genome was detected; all other differences could be explained by resolution of mapping techniques used. Fourteen of the most informative microsatellite markers in the first 27 cM of the map were genotyped across the entire MARC swine resource population, increasing the number of markers typed from 2 to 14 and more than doubling the number ofgenotyped animals with ovulation rate data (295 to 600). Results from the revised data set for the QTL analysis, assuming breed specific QTL alleles, indicated that the most likely position of the QTL resided at 4.85 cM on the new linkage map (F1,592 = 20.5150, genome-wide probability less than 0.015). The updated estimate of the effect of an allele substitution was -1.65 ova for the Meishan allele. The F-ratio peak was closest to markers for MAN2B2 (4.80 cM) and was flanked on the other side by markers for PPP2R2C. Two positional candidate genes included in this study are MAN2B2 and RGS12. These results validate the presence of a QTL affecting ovulation rate on chromosome 8 and facilitate selection of positional candidate genes to be evaluated.     

Effects of porcine leptin receptor gene polymorphisms on backfat thickness,fat area ratios by image analysis,and serum leptin concentrations in a Duroc purebred population

The leptin receptor (LEPR) gene is considered a candidate gene for fatness traits. It is located on SSC 6 in a region in which quantitative trait loci (QTLs) for backfat thickness (BF), fat area ratios, and serum leptin concentration (LEPC) have previously been detected in a Duroc purebred population. The objectives of the present study were to identify porcine LEPR polymorphisms and examine the effects of LEPR polymorphisms on fatness traits in this same population. The Duroc pigs (226 to 953 pigs) were evaluated for BF, fat area ratios using image analysis, and LEPC. A total of seven single nucleotide polymorphisms (SNPs) in the full‐length LEPR coding region were identified in pigs from the base population. Four non‐synonymous SNPs of the LEPR gene and 15 microsatellite markers on SSC 6 were then genotyped in all pigs. During candidate gene analysis, we detected significant effects of the non‐synonymous SNP c.2002C>T in exon 14 on all traits. In fine mapping analysis, significant QTLs for BF, fat area ratios, and LEPC were detected near the LEPR gene in the same region. These results indicated that the c.2002C>T SNP of LEPR has a strong effect on BF, fat area ratios and LEPC.