大白猪和长白猪EPOR基因第4内含子多态性与繁殖性状的关联分析

本研究以大白猪和长白猪为研究对象,以促红细胞生成素受体(erythropoietin receptor,EPOR)基因作为产仔性状的候选基因,采用PCR-SSCP方法来检测EPOR基因第4内含子C\T突变多态性,研究此位点多态性与猪繁殖性状之间的关联性。结果发现,在长白猪和大白猪中存在AA、AB和BB共3种基因型,在2个群体中处于中度多态。利用SAS 8.0软件采用最小二乘法拟合线性模型,将不同基因型与总产仔数(TNB)、产活仔数(NBA)和初生重(WB)进行了关联分析,结果表明,长白猪初产母猪BB基因型个体的WB显著高于AB、AA基因型个体(P<0.05);经产母猪BB基因型个体的TNB显著高于AB基因型个体(P<0.05);B等位基因对初产母猪TNB、NBA和WB均表现为正效应。大白猪初产母猪BB基因型个体的TNB、NBA和WB都高于AA型和AB型个体,但差异均不显著(P>0.05);经产母猪AB基因型个体的WB显著高于AA基因型个体(P<0.05);B等位基因对初产母猪TNB、NBA和WB都表现为正效应。     

猪VEGF基因5′调控区多态性与产仔数的关联分析

研究通过PCR-SSCP技术对大白猪、长白猪和杜洛克猪共计574头母猪的VEGF基因进行多态分析,并将基因多态性与猪总产仔数(TNB)和产活仔数(NBA)进行关联分析,旨在探讨猪VEGF基因多态性对其产仔数的影响。结果表明:在VEGF 5′调控区内有2个突变位点。P1产物第191位点处由A突变为G,在3个猪品种中只发现AA、AG 2种基因型,其中AG为优势基因型,A为优势等位基因。长白猪第1胎中AG型的TNB和NBA比AA型分别高出1.34头和1.44头(P<0.01),杜洛克猪所有胎次中AG与AA的TNB差异显著(P<0.05),NBA差异极显著(P<0.01)。P2产物第103位点处由C突变为T,在3个猪品种中发现TT、TC、CC 3种基因型,T为优势等位基因。TT、TC分别比CC型的TNB高出2.63头(P<0.05),在杜洛克所有胎次中TC比TT的TNB和NBA分别高出0.84头和0.94头(P<0.05)。P1位点和P2位点能否作为产仔性状的标记基因还需进一步研究。     

Evaluation of FABP2 as candidate gene for a fatty acid composition QTL in porcine chromosome 8

The objective of this work was to analyse the porcine Fatty acid binding protein 2, intestinal ( FABP2 ) gene as a candidate gene for a fatty acid composition quantitative trait loci (QTL) previously described on porcine chromosome 8 in an Iberian by Landrace F₂ cross (IBMAP). Re-sequencing of the porcine FABP2 gene in three Iberian and eight Landrace parental animals resulted in the identification of three single-nucleotide polymorphisms, all of them localized in intron 1. The polymorphism FABP2 :g.412T>C, localized in intron 1, and two additional microsatellites were genotyped in the IBMAP population in order to perform an association test of the FABP2 gene and to better define the QTL position previously described. Association analyses of the FABP2 :g.412T>C with the fatty acid composition traits were not significant in simple association and marker-assisted association tests, suggesting that the FABP2 region sequenced is not responsible for the QTL. However, the addition of three new markers to the pedigree allowed us to define the S0144–SW61 marker interval as the most likely QTL position, facilitating the future study of other candidate genes for this QTL.     

A new HpaII PCR-RFLP within the porcine prolactin receptor (PRLR) gene and study of its effect on litter size and number of teats

DNA polymorphism of the porcine prolactin receptor gene ( PRLR ) was investigated and used to study its effect on litter size and number of teats in pigs. By means of PRLR gene sequence homology in pig, human and other species, primers were designed for PCR amplification within 5' unknown (to date) part of the prolactin receptor gene in pigs. In this part of the gene, a new polymorphism with Hpa II restriction endonuclease was detected. Alu I polymorphism described before and our new Hpa II polymorphism were used to study the associations with reproduction traits. The PCR restriction fragment length polymorphism (PCR-RFLP) method was used to genotype Alu I and Hpa II loci of the PRLR gene in line A with 83 sows of Landrace breed and in two lines (B and C) with 75 and 86 Large White sows, respectively. Statistical analysis of 1020 litters showed that Alu I locus was associated with litter size mainly in Landrace and affected the first parities, while Hpa II locus of the gene was associated with the same traits in Landrace and Large White pigs and mainly affected numbers of weaned of pigs. The magnitude of the effect varied by population with the effects exceeding two pigs per litter in Landrace line and 1 pig per litter in Large White populations.     

4猪种Nramp1基因第6内含子多态性研究

天然抗性巨噬蛋白(Nramp)基因是与人、鼠的一些病原微生物的易感性和抗性有关的重要候选基因。为了研究猪Nramp1基因的多态性,利用PCR-RFLP技术检测了杜洛克、大白猪、长白猪和合作猪共270头个体Nramp1基因第6内含子NdeⅠ酶切位点多态性。结果表明,4个猪种群共检测到3种基因型(AA、AB和BB),其中AB基因型为杜洛克、大白猪和长白猪的优势基因型;AA基因型为合作猪的优势基因型。经卡方适合性检测,杜洛克、合作猪和长白猪处于Hardy-Weinberg平衡状态(P>0.05),大白猪处于Hardy-Weinberg不平衡状态。多态信息含量分析显示,Nramp1基因的第6内含子NdeⅠ酶切位点在各猪种表现出中度多态性。     

部分引进猪种α1-岩藻糖转移酶基因多态性研究

本研究采用PCR RFLP技术对4个引进猪种杜洛克、约克夏、长白猪和皮特兰共计122头猪的FUT1基因进行了多态性分析。结果表明：4个猪种在该FUT1基因座位存在多态性，但抗性基因型AA仅在杜洛克和皮特兰猪中分布。卡方检验结果表明，长白猪与皮特兰间FUT1基因基因型分布差异极显著，长白猪与杜洛克猪之间差异显著，约克夏与皮特兰间差异也显著。     

鸡脂滴包被蛋白基因内含子5多态性与胴体及脂肪性状的相关性分析

本试验采用PCR-RFLP方法检测脂滴包被蛋白基因(perilipin,PLIN)在济宁百日鸡、汶上芦花鸡等5个地方鸡种和1个培育品系中的遗传多态性,并分析了多态位点不同基因型与鸡胴体及脂肪性状的相关性。结果发现,在6个供试群体中检测到1个Pst Ⅰ酶切突变位点,测序证实为新发现的鸡PLIN基因2224 bp处G→T突变,该突变位点在供试群体中共检测到两种基因型(A1A1和A1A2),等位基因A1在所有供试群体中均表现为优势等位基因。关联分析结果表明,PLIN基因2224多态位点对鸡胴体性状和脂肪性状影响均不显著(P＞0.05)。多重比较结果表明,A1A1基因型个体在胴体和脂肪性状上总体优于A1A2基因型个体,但各性状均值在两种基因型个体间差异均不显著(P＞0.05)。本研究结果表明,该位点不能作为鸡胴体和脂肪性状的有效分子标记。     

Investigation of two candidate genes for meat quality traits in a quantitative trait locus region on SSC6: the porcine short heterodimer partner and heart fatty acid binding protein genes

A highly significant quantitative trait locus (QTL) on pig chromosome 6, affecting intramuscular fat (IMF), has previously been detected by our group and others. Two genes of positional and biological interest, the small heterodimer partner (SHP; NR0B2) and the heart fatty acid binding protein (FABP3; H‐FABP), were investigated for meat quality traits and IMF respectively. SHP was partially sequenced (GenBank: DQ002896 and DQ002897 ) and mapped to the QTL region on porcine chromosome 6, affecting IMF. The map shows no recombination between SHP and FABP3, which was previously mapped to the same QTL region. Twelve single nucleotide polymorphisms were detected in the sequenced region of SHP gene. Haplotype information was used to investigate association between genetic variation and different meat quality traits. SHP haplotype combinations were found to have significant effect on connective tissue. However, further studies are needed to evaluate this possible association more effectively. The FABP3 is involved in fatty acid transport and has been studied as a candidate gene for IMF by several research groups. In our study, FABP3 genotypes were confirmed to be significantly associated with IMF in pigs. The average content of IMF in our population was 1.6%, which may indicate that the FABP3 polymorphism explains as much as 30–35% of the variation in IMF in our pig cross‐population.     

Intramuscular fat content and genetic variants at fatty acid-binding protein loci in Austrian pigs.

Intramuscular fat is an important meat quality trait in pig production. Previously, genetic variants of the heart fatty acid-binding protein (H-FABP) gene and the adipocyte fatty acid-binding protein (A-FABP) gene were suggested to be associated with intramuscular fat content. The objective of this investigation was to study these associations in the three most important Austrian breeding populations (Piétrain, Large White, and Landrace). Restriction fragment length polymorphism analysis of the H-FABP gene revealed a new MspI polymorphic site and genetic variation in all three breeds. Microsatellite analysis of the A-FABP locus showed up to nine different microsatellite alleles segregating. In Austrian breeds, no significant influence of the A-FABP and H-FABP gene polymorphisms on intramuscular fat could be detected. We also evaluated possible associations between the genetic variations at the H-FABP and A-FABP loci and other growth and carcass traits (average daily gain, feed conversion ratio, lean meat content, pH values, meat color, and drip loss). With regard to the extent of the effects, these genetic markers cannot be recommended for selection on growth and carcass traits in Austrian breeding populations.     

延黄牛ALDH1A1基因内含子4多态性及其与肉用性状的相关性分析

试验旨在研究乙醛脱氢酶1A1（acetaldehyde dehydrogenase 1A1，ALDH1A1）基因内含子4多态性及其对延黄牛肉用性状的影响。选取99头18月龄延黄牛母牛，采用PCR扩增产物Sanger直接测序法测定99头延黄牛ALDH1A1基因内含子4的单核苷酸多态性（SNP），运用SPSS 19.0软件分析ALDH1A1基因内含子4 SNP与延黄牛肉用性状（体高、体斜长、十字部高、胸围、腹围、管围、体重、背膘厚、眼肌面积和肌内脂肪）的关联性。结果显示，延黄牛ALDH1A1基因内含子4存在C/T突变位点，共发现3种基因型：TT、TC和CC，2种等位基因：T和C，其中TC为优势等位基因型，C为优势等位基因。卡方适合性检验结果发现，该SNP在延黄牛群体中处于Hardy-Weinberg平衡状态（P>0.05）。群体遗传参数分析发现，该突变位点的杂合度（He）相对较低，表明其在延黄牛群体中的变异较小，该位点属于中度多态（0.250.05）。结果表明，ALDH1A1基因内含子4在延黄牛中存在突变，能否作为延黄牛肉用性状的遗传标记有待于扩大样本数量进一步研究。     

不同猪种TGF-β1基因单核苷酸多态性分析

以二花脸猪、大白猪、长白猪、皮特兰猪和圣特西猪共计861 头为研究材料,采用PCR-SSCP技术,对猪TGF β1基因6 7外显子区的1156 bp序列进行多态性分析,发现一个多态位点。经克隆测序分析,位于第6内含子区内存在C→T突变,该突变位点为第7外显子上游的第9位碱基(序列:AJ621785中的第1043位点)。对不同猪群的基因型和基因频率统计结果表明,二花脸猪以等位基因T为主,而大白猪、长白猪、皮特兰猪和圣特西猪则以等位基因C为主,且各猪群均处于Hardy Weinberg平衡。     

Candidate gene markers for litter size in different German pig lines.

Three diallelic RFLP markers at candidate gene loci for litter size, the estrogen receptor (ESR) gene, the prolactin receptor (PRLR) gene, and the retinol-binding protein 4 (RBP4) gene, were evaluated for their association with the number of piglets born alive in different German pig lines. Genotyping was performed on boars and sows belonging to three different genetic groups from a single farm. Information on 8,336 litter records from 2,159 sows (German Landrace, n = 1,672; Duroc, n = 214; and a synthetic line, n = 273) was used in the analyses with respect to litter size. Growth performance traits were only analyzed for the synthetic line. The ESR locus showed no polymorphism in the tested boars of the German Landrace and Duroc lines. In the synthetic line, the frequency for the A allele was 0.90 and no homozygous BB animal was detected. No significant associations of ESR alleles with number of piglets born alive, backfat thickness, or average daily gain were observed. A new PCR-RFLP was developed for testing the PRLR polymorphism. The frequencies of PRLR allele A were 0.40 in the German Landrace, 0.49 in the synthetic, and 0.82 in the Duroc line. In the Duroc line, a small additive effect of the allele B on litter size was observed. The allelic substitution effect was 0.71 piglets born alive across all parities (P = 0.05). No significant associations of the PRLR locus with litter and growth performance traits were detected. The frequencies of RBP4 allele A ranged from 0.62 in the synthetic line to 0.67 in the German Landrace to 0.85 in the Duroc line. For the genotyped sows of the synthetic line, there was no indication of a favorable effect of the A allele with respect to litter size. Results of this study demonstrate that allele effects differ between lines or populations. This may be due to possible different linkage phases between the marker alleles and the causal mutations in the different lines. The results may also be explained by many minor genes affecting litter size. A selection strategy should be designed for each line separately and should always consider possible pleiotropic effects.     

大白猪和长白猪EPOR基因第4内含子多态性与繁殖性状的关联分析

本研究以大白猪和长白猪为研究对象,以促红细胞生成素受体（erythropoietin receptor,EPOR）基因作为产仔性状的候选基因,采用PCR-SSCP方法来检测EPOR基因第4内含子C＼T突变多态性,研究此位点多态性与猪繁殖性状之间的关联性。结果发现,在长白猪和大白猪中存在AA、AB和BB共3种基因型,在2个群体中处于中度多态。利用SAS 8.0软件采用最小二乘法拟合线性模型,将不同基因型与总产仔数（TNB）、产活仔数（NBA）和初生重（WB）进行了关联分析,结果表明,长白猪初产母猪BB基因型个体的WB显著高于AB、AA基因型个体（P＜0.05）;经产母猪BB基因型个体的TNB显著高于AB基因型个体（P＜0.05）;B等位基因对初产母猪TNB、NBA和WB均表现为正效应。大白猪初产母猪BB基因型个体的TNB、NBA和WB都高于AA型和AB型个体,但差异均不显著（P＞0.05）;经产母猪AB基因型个体的WB显著高于AA基因型个体（P＜0.05）;B等位基因对初产母猪TNB、NBA和WB都表现为正效应。     

京海黄鸡LYZ基因外显子的遗传多态性及其与体重的关联分析

试验旨在研究鸡溶菌酶基因（LYZ）对鸡体重的影响。以200只京海黄鸡为研究群体,采用PCR-SSCP和DNA测序的方法检测LYZ基因单核苷酸多态性,并将所发现的SNPs与体重进行关联分析。结果发现,共3个突变位点,且均为沉默突变,未引起氨基酸序列的改变;经χ2适合性检验,除1411位点外,其他2个突变位点均显著偏离Hardy-Weinberg平衡;111位点AA基因型个体12和16周龄体重显著高于GG基因型个体（P＜0.05）,1411位点AA和GA基因型个体各周龄体重均高于GG基因型个体,且4周龄体重差异达到了显著水平（P＜0.05）,1426位点各基因型对体重没有显著影响（P＞0.05）。单倍型分析结果显示,除4周龄外,ANC个体各周龄体重均较大,GAC个体4周龄体重较大,这与上述各位点基因型对体重影响的分析结果相一致,暗示在选种、育种过程中,可将鸡LYZ基因作为京海黄鸡体重候选基因应用于遗传标记辅助选择。     

鸭THRSPα基因内含子多态与生长和屠体性状相关性研究

根据克隆出的鸭THRSPα基因全长序列,采用PCR-RFLP法检测THRSPα内含子多态与238只6周龄北京鸭生长、屠体性状的关系。结果发现:在THRSPα内含子第239处存在C-A碱基突变,该突变为SacI酶切位点,C碱基基因频率占95%,为优势等位基因,χ2检验结果表明,该群体处于Hardy-Weinberg平衡状态。最小二乘分析表明:THRSPα内含子SacI位点获得的2种基因型CC和CA与6周龄体重、屠宰性状的关系中,除CC型肝脏率极显著(P<0.01)低于CA型,其他性状两基因型间的差异均不显著(P>0.05)。     

黔北麻羊FABP3基因多态性及其与生长性状的相关性研究

旨在探究脂肪酸结合蛋白(FABP3)基因在黔北麻羊中的遗传多态性,进一步揭示FABP3基因与黔北麻羊生长性状间的关联性,为今后运用分子标记辅助育种方法提高黔北麻羊的生长性状提供理论依据。本研究以120只6月龄黔北麻羊为试验对象,采用PCR产物直接测序技术检测FABP3基因全部外显子区的遗传多态性,并与体重、体高、体斜长、胸围、管围等生长性状进行关联性分析。结果:在黔北麻羊FABP3基因第5外显子和3′非翻译区筛选出2个SNPs位点,分别为g.13665051C>T和g.13664737G>A,其中g.13665051C>T位点为同义突变,产生3种基因型:CC、CT和TT,g.13664737G>A位点产生3种基因型:GG、AG和AA,卡方(χ²)检验结果显示,这2个SNPs位点处于中度多态且在黔北麻羊群体中均未偏离Hardy-Weinberg平衡(P>0.05)。关联分析显示,g.13665051C>T位点中CT基因型个体的体高、体斜长、胸宽和管围显著高于CC和TT基因型个体(P<0.05);CT基因型个体的胸深、胸围显著...     

猪脂肪细胞型脂肪酸结合蛋白基因BsmI位点多态性与肌内脂肪含量的相关研究

本研究利用PCR-RFLPs标记技术,对中国地方猪种雅南猪、大河猪、培育品种大河乌猪以及三元杂交群体DLY共113头个体的脂肪细胞型脂肪酸结合蛋白基因的内含子Ⅰ区域进行了分析研究,发现BsmΙ酶切位点具有多态性,并分为3种基因型(即AA、BB和AB型),通过构建固定效应模型,分析了基因型与肌内脂肪含量的关系。结果表明:基因型AA的肌内脂肪含量极显著的高于基因型AB和BB(P<0.01)。     

梅花鹿MSRB3基因多态性及其与茸重性状的关联分析

【目的】本研究旨在探索梅花鹿甲硫氨酸亚砜还原酶B3(methionine sulfoxide reductase B3,MSRB3)基因多态性及其与茸重性状的关联性。【方法】选取高、低产茸量的梅花鹿各8只,应用DNA直接测序法检测基因变异位点。采用MassARRAY~? SNP分型技术对314头24月龄梅花鹿MSRB3基因进行基因分型,并结合梅花鹿茸重性状数据进行了关联分析和单倍型分析。【结果】在梅花鹿MSRB3基因中共发现6个SNPs位点,其中2个SNPs位点位于外显子区域,且突变均未引起氨基酸改变,属于同义突变,其余4个SNPs位点均存在于内含子区域。分型结果显示4个样本未分型成功,其余310个样本进行后续分析。各位点观测杂合度和期望杂合度基本一致,g.44455582 T>C、g.44455759 C>T、g.44414424 T>C、g.44350306 T>C及g.44340836 G>A等5个位点的杂合度较高,均属于中度多态(0.25C位点杂合度较低,属于低度多态(PIC<0...     

Characterization of polymorphisms of transferrin receptor and their association with susceptibility to ETEC F4ab/ac in pigs

Diarrhoea caused by enterotoxigenic Escherichia coli (ETEC) expressing F4 (F4ab, F4ac and F4ad) fimbriae is a significant cause of mortality and morbidity in newborn and weaned pigs. The locus controlling susceptibility towards ETEC F4ab/ac has been mapped to SSC13q41, in which TFRC (transferrin receptor) was localized and considered as a positional candidate gene for ETEC F4ab/ac receptor. In this study, we determined susceptibility/resistance to ETEC F4ab/ac in a total of 755 F2 animals from a White Duroc x Erhualian intercross using a microscopic enterocyte adhesion assay. We identified two TFRC polymorphisms (SNPs 591 A>G and 632 A>G) in a single exon after comparative sequencing analysis of 2371-bp amplicons containing the complete coding region of TFRC using RNA of eight full-sib F2 animals with susceptible and resistant phenotypes. The intron sequences flanking the two exon polymorphisms were obtained, revealing an intron polymorphism (SNP 291 C>T). We genotyped the 19 founder animals of the White Duroc x Erhualian intercross for the identified polymorphisms, showing that only the 291 C>T polymorphism is a highly informative marker. We further genotyped all 59 F1 and 755 F2 animals for the 291 C>T polymorphism, and the association of this polymorphism with susceptibility/resistance to ETEC F4ab/ac in these F2 animals was evaluated by the transmission disequilibrium test. The result showed that the 291 C>T polymorphism is not a causal mutation, however, has a significant linkage disequilibrium with the ETEC F4ab/ac, especially F4ac receptor locus.     

摩拉水牛和尼里-拉菲水牛PRKAA2基因SNPs检测及遗传多样性分析

为研究水牛蛋白激酶AMP活化的催化亚基α2（protein kinase AMP-activated catalytic subunit alpha 2,PRKAA2）基因多态性,本试验以摩拉水牛和尼里-拉菲水牛基因组DNA为模板,扩增PRKAA2基因外显子4及内含子3部分序列,通过常规测序法检测其SNP并进行遗传多样性分析。结果发现,PRKAA2基因外显子4内存在1个SNP位点（c.462 G>A）,PRKAA2基因内含子3部分序列存在3个SNPs位点（IVS3.557 T>C、IVS3.560 C>T和IVS3.565 G>A）。经遗传多样性分析表明,在c.462 G>A位点的野生纯合型和杂合型比突变纯合型更有优势,IVS3.557 T>C和IVS3.560 C>T位点的突变纯合型为非优势基因型,IVS3.565 G>A位点杂合型为优势基因型。IVS3.565 G>A位点在摩拉水牛群体中处于Hardy-Weinberg非平衡状态;c.462 G>A位点在尼里-拉菲水牛群体中处于Hardy-Weinberg非平衡状态。4个SNPs位点在摩拉水牛群体中均为中度多态;c.462 G>A、IVS3.557 T>C位点在尼里-拉菲水牛群体中为低度多态,IVS3.560 C>T、IVS3.565 G>A位点为中度多态。IVS3.557 T>C位点在两个水牛群体中杂合度较低。说明摩拉水牛IVS3.565 G>A位点和尼里-拉菲水牛c.462 G>A位点的基因型频率和基因频率遗传状态不平衡,尼里-拉菲水牛群体中IVS3.557 T>C位点遗传变异小,选择潜力不高。4个多态位点可以构建5种单倍型,其中T-C-G-G是摩拉水牛群体和尼里-拉菲水牛群体的优势单倍型。综上,本研究检测的摩拉水牛和尼里-拉菲水牛PRKAA2基因上4个SNPs位点可为水牛标记辅助选择育种提供参考。