猪PRLR基因PCR—SSCP多态性与产仔性能的关联分析

采用PCR-SSCP技术分析了杜洛克猪、长白猪、大白猪、马身猪、山西黑猪和山西白猪等6个品种472个个体催乳素受体基因(PRLR)的多态性,检测到A、B、C 3个等位基因和AA、AB、AC、BB、CC 5种基因型.在马身猪中, B等位基因为优势基因,频率为0.55;其他品种中,优势基因为A等位基因,频率分布在0.79～0.89之间;C等位基因除在大白猪频率略高外(0.20),在其他品种中频率都很低,在0～0.09之间.对AA、BB、CC三种纯合子进行克隆测序和同源序列比较,发现在扩增片段内有6处SNP,都发生在PRLR基因的第8内含子,分别是内含子8第26位、54位和99位的C→T突变,47位和68位的A→G突变,63位的G→A突变.利用最小二乘分析研究了PRLR基因型对母猪头胎总产仔数和产活仔数的影响,结果表明PRLR基因不同基因型母猪的头胎总产仔数和产活仔数差异均不显著.     

猪H-FABP基因PCR-SSCP分析

应用PCR-SSCP方法分析了H-FABP基因在山西白猪、马身猪、大白猪、长白猪和杜洛克猪5个猪种的多态性。结果表明:在H-FABP基因内含子1中发现了多态位点,该位点上具有两个等位基因A和B,马身猪BB基因型频率最高,B等位基因频率明显高于A等位基因频率;其余4个猪种AA基因型频率最高。序列测定的结果表明,SSCP的变异是由碱基C→T的替换造成的。     

西部地区主要猪种和野猪H-FABP基因分子标记

利用PCR-RFLP(Hinf I、Hae Ⅲ和Msp I 3种限制性内切酶)分子标记技术,检测了杜洛克猪、长白猪、大白猪、内江猪、荣昌猪、汉江黑猪、汉中白猪、八眉猪和野猪共计265头猪H-FABP基因5′-上游区和第二内含子区的遗传变异,并利用最小二乘模型分析了H-FABP基因对猪肌内脂肪含量的遗传效应。结果表明：（1）在Hinf I-RFLP位点上,上述品种和野猪均存在多态性,其中大白猪、八眉猪、汉江黑猪、汉中白猪和野猪表现为低度多态,杜洛克、长白猪、内江猪和荣昌猪为中度多态;除汉江黑猪（P<0.05）和野猪（P<0.01）外,其他猪种基因频率和基因型频率都处于Hardy-Weinderg平衡状态（P>0.05）;而在Hae Ⅲ-RFLP和Msp I-RFLP位点上,仅内江猪、荣昌猪、汉江黑猪和八眉猪为单态;（2）9种基因型对肌内脂肪（IMF）含量的影响,HH>Hh>hh,DD相似文献   

猪MyoG基因3''''端PCR-SSCP遗传多态性及其遗传效应

采用PCR—SSCP方法对长白猪、大白猪、杜洛克猪、山西黑猪和马身猪共636头猪的肌细胞生成素（Myogenin,简称MyoG）基因3’端的遗传多态性进行检测,分析MyoG基因对猪的初生体质量、断奶体质量、6月龄体质量和背膘厚的影响。根据已发表的猪MyoG基因3’端侧翼序列设计3对引物,发现F1／R1引物对扩增的片段有多态性。统计结果发现：长白、大白、杜洛克猪种B基因为优势基因,其基因频率分别为0．8807、0．7256和0．8581：山西黑猪种4基因为优势基因,其基因频率为0．9359;马身猪种只检测到4基因。χ^2独立性检验表明,基因型分布在外来猪种（长白猪、大白猪、杜洛克猪）与地方猪种（山西黑猪、马身猪）间存在极显著差异（P〈0．01）。固定效应模型分析结果表明,初生体质量基因型闻差异显著（P〈0．05）,而断奶体质量、6月龄体质量和背膘厚基因型闻差异不显著（P〉0．05）。最小二乘分析结果表明,BB基因型与其它2种基因型比较有较大的初生质量,同似和AB型比较差异极显著（P〈0．01）。因此,推测MyoG基因对个体的初生体质量存在一足的影响,选择带有B等位基因的个体有望提高个体的初生体质量。     

不同品种猪肌肉生长抑制素基因单核苷酸多态性分析

用PCR-RFLPs和PCR-SSCP分析方法,对"双肌臀”大白猪、大白猪、长白猪、杜洛克、汉普夏、皮特兰、二花脸、东北民猪、湖北白猪和部分杂交猪等不同品种猪肌肉生长抑制素基因3＇编码区、5＇调控区及内含子1区3个单核苷酸多态性位点(SNPs)进行了分析.结果表明,3＇编码区的SNP发生的频率较低,在274头猪中未检出突变纯合体.对5＇调控区的SNP,引进猪种(大白猪、长白猪、杜洛克、汉普夏和皮特兰)及其杂交猪以等位基因T为主,二花脸和湖北白猪则以等位基因A为主,均偏离Hardy-Weinberg平衡状态(P＜0.01).东北民猪的3种基因型近乎相等,处于Hardy-Weinberg衡状态.对内含子1区的SNP,大白猪及其与长白猪的杂交猪等位基因G占优势,二花脸和湖北白猪则以等位基因A为主,均偏离Hardy-Weinberg平衡状态(P＜0.01);东北民猪和大二猪的等位基因G和A近乎相等,处于Hardy-Weinberg平衡状态."双肌臀”大白猪在5＇调控区和内含子1区这两个位点的A等位基因稍高于普通大白猪.5＇调控区和内含子1区SNPs所产生的等位基因表现出连锁遗传现象.     

猪MyoG基因3′端PCR-SSCP遗传多态性及其遗传效应(英文)

采用PCR-SSCP方法对长白猪、大白猪、杜洛克猪、山西黑猪和马身猪共636头猪的肌细胞生成素(Myogenin,简称MyoG)基因3′端的遗传多态性进行检测,分析MyoG基因对猪的初生体质量、断奶体质量、6月龄体质量和背膘厚的影响。根据已发表的猪MyoG基因3′端侧翼序列设计3对引物,发现F1/R1引物对扩增的片段有多态性。统计结果发现:长白、大白、杜洛克猪种B基因为优势基因,其基因频率分别为0.8807、0.7256和0.8581;山西黑猪种A基因为优势基因,其基因频率为0.9359;马身猪种只检测到A基因。χ2独立性检验表明,基因型分布在外来猪种(长白猪、大白猪、杜洛克猪)与地方猪种(山西黑猪、马身猪)间存在极显著差异(P<0.01)。固定效应模型分析结果表明,初生体质量基因型间差异显著(P<0.05),而断奶体质量、6月龄体质量和背膘厚基因型间差异不显著(P>0.05)。最小二乘分析结果表明,BB基因型与其它2种基因型比较有较大的初生质量,同AA和AB型比较差异极显著(P<0.01)。因此,推测MyoG基因对个体的初生体质量存在一定的影响,选择带有B等位基因的个体有望提高个体的初生体质量。     

猪Kappa阿片受体基因外显子部分序列单核苷酸多态性分析

应用PCR SSCP的方法对大白猪、长白猪和杜洛克猪的Kappa阿片受体 (Kappaopioidreceptor,简称KOR)基因进行单核苷酸多态性检测和分析 ,研究Kappa阿片受体基因作为候选基因 ,影响母猪行为规癖性状的可能性。根据Kappa阿片受体基因外显子的部分序列设计 3对引物 ,发现F1/R1引物对扩增的片段有多态性。对两种纯合子片段克隆并测序表明 ,mRNA第 10 5处有一C→T的单碱基突变 ,为沉默突变。统计结果发现 ,3种基因型 (AA、AB、BB)在各品种中的分布不一致 ,χ2 独立性检验差异极显著 (P <0 0 1)。将 3种基因型同行为规癖性状进行统计分析 ,结果表明BB基因型与其他两种基因型相比表现较高的静止站立行为 ,同AA和AB型比较差异极显著 (P<0 0 1) ,其他 4种行为性状基因型间差异不显著。因此 ,推测Kappa阿片受体基因SNP对母猪静止站立行为性状存在一定的影响     

猪CAST基因的遗传多态性及遗传效应分析

采用PCR—SSCP方法对猪cAST基因遗传多态性进行分析。并研究基因型与肉质性状和背膘厚的相关性。根据猪CAST基因的cDNA序列（M20160）设计7对引物。结果在F1／R1。F6／R6引物对扩增的片段上发现了多态性。并对纯合子进行测序。发现317位A—G突变。2042位G—C突变。基因型在不同猪种分布的多重比较结果表明。长白猪、大白猪和杜洛克猪与沂蒙黑猪和莱芜猪比较差异极显著（P〈0．01）。固定效应模型分析结果表明,嫩度及背膘厚基因型间差异显著（P〈0．05）,而pH值、温度及滴水损失基因型间差异不显著（P〉0．05）。最小二乘分析结果表明,不同猪种比较,屠宰12h和24h后肌肉温度、30min和1h后pH值及滴水损失差异显著（P〈0．05）;BBCC和BBDD单元型个体与其他单元型个体比较肌肉嫩度的差异显著（P〈0．01）。AACC和AADD单元型个体与其他单元型个体比较背膘厚的差异显著（P〈0．01）。因此,推测CAST基因对猪肉品质及背膘厚存在一定的影响。将CAST基因应用于猪育种过程中的标记辅助选择将可以改善猪肉品质。加快猪的育种进程。     

大白猪BF基因多态性与繁殖性状及胎盘效率关联研究

文章以BF基因为产仔数的候选基因, 运用PCR-RFLP方法对大白猪BF基因内含子1序列进行分析, 发现内含子1第79 bp处发生了C→T突变, 命名为BF-intron 1-C79T。所扩增PCR产物经SmaⅠ酶切后, 可分出3种基因型, 分别为CC、CT和TT。经χ²适合性检验, 大白猪群体在该位点达到了Hardy-Weinberg平衡状态。将不同基因型与总产仔数(TNB)、产活仔数(NBA)、初生重(BW)和胎盘效率(PE)进行关联分析, 结果发现, 初产母猪在该位点上, CC基因型比CT基因型个体的TNB和NBA均多3.10头, 但差异不显著(P>0.05); 在BW和PE上, CC基因型均高于CT基因型, 差异不显著(P>0.05)。经产母猪在该位点上, CC基因型比TT基因型个体的TNB和NBA分别多3.45头和3.92头, PE高23.80%, 均达到显著水平; 但CT基因型个体与CC和TT个体之间的TNB 、NBA和PE差异不显著(P>0.05)。因此, BF-intron 1-C79T位点可作为繁殖性状及胎盘效率的潜在分子育种标记, 具有很大的研究价值。     

猪MyoG基因内含子1的一个遗传多态性及其遗传效应分析

以长白猪(73),大白猪(68),沂蒙黑猪(57)和莱芜猪(83)为研究对象,采用PCR-SSCP方法 对猪MyoG基因遗传多态性进行分析,并研究基因型与初生重、日增重、肌肉嫩度和背膘厚的相关性.根据猪MyoG 基因的DNA序列（M14331）设计引物,结果在内含子1扩增的片段上发现了一个多态性,检测到2个等位基因(A、B),3种基因型（AA、AB、BB）,并对纯合子进行测序,发现2　943位G→C突变.基因型在不同猪种分布的多重比较,结果表明,大白猪与长白猪、沂蒙黑猪和莱芜猪比较差异显著（P<0.05）,莱芜猪与长白猪和沂蒙黑猪比较差异显著（P<0.05）,长白猪与沂蒙黑猪比较差异不显著（P>0.05）.固定效应模型分析结果表明,初生重及嫩度基因型间差异显著（P<0.01）,而日增重及背膘厚基因型间差异不显著（P>0.05）.最小二乘分析结果表明,BB基因型个体与AA基因型个体比较肌肉嫩度的差异显著（P<0.05）,AA基因型个体与AB和BB基因型个体比较背膘厚的差异显著（P<0.05）.因此,推测MyoG基因对猪肉品质、生长速度及背膘厚存在一定的影响,将 MyoG基因应用于猪育种过程中的标记辅助选择将可以改善猪肉品质,提高生长速度,加快猪的育种进程.     

Unexpectedly high allelic diversity at the KIT locus causing dominant white color in the domestic pig.

Mutations in KIT encoding the mast/stem cell growth factor receptor (MGF) are responsible for coat color variation in domestic pigs. The dominant white phenotype is caused by two mutations, a gene duplication and a splice mutation in one of the copies leading to skipping of exon 17. Here we applied minisequencing and pyrosequencing for quantitative analysis of the number of copies with the splice form. An unexpectedly high genetic diversity was revealed in white pigs. We found four different KIT alleles in a small sample of eight Large White females used as founder animals in a wild boar intercross. A similar number of KIT alleles was found in commercial populations of white Landrace and Large White pigs. We provide evidence for at least two new KIT alleles in pigs, both with a triplication of the gene. The results imply that KIT alleles with the duplication are genetically unstable and new alleles are most likely generated by unequal crossing over. This study provides an improved method for genotyping the complicated Dominant white/KIT locus in pigs. The results also suggest that some alleles may be associated with negative pleiotropic effects on other traits.     

Genetic Variation and Association of Insulin-Like Growth Factor Binding Protein-3 with Performance in Swine

Genetic variation of insulin-like growth factor binding protein-3 was analyzed in 17 pig breeds (14 native Chinese and 3 European). Using PCR-single strand conformation polymorphism, we found a polymorphism in intron 2, and this SNP was the combined mutation of G897T-G903A-C911T. The Chinese breeds carried a higher TAT/TAT genotype frequency (over 50%), except for Bamei (22%), Yujiang Black (0.0%), and Erhualian (10.0%); the European breeds had a higher GGC/GGC genotype frequency (Large White 1.67%, Landrace 13.89%, Duroc 0.0%). The allelic frequency of TAT in Chinese breeds was over 50%, except for Yujiang Black (12.5%); the allelic frequency of GGC was over 50% in all European breeds. The effect of genotype on 43 performance traits was investigated in one population (Lantang × Landrace). Pigs with the TAT/TAT genotype had higher B-point and C-point back-fat thickness than pigs with the GGC/GGC genotype. The TAT/TAT pigs also scored higher in meat color than the GGC/GGC pigs. These results implied that IGFBP-3 may affect meat quality and carcass traits.     

Chinese white Rongchang pig does not have the dominant white allele of KIT but has the dominant black allele of MC1R

The mast/stem cell growth factor receptor (KIT) and melanocortin receptor 1 (MC1R) mutations are responsible for coat color phenotypes in domestic pigs. Rongchang is a Chinese indigenous pig breed with a white coat color phenotype. To investigate the genetic variability of the KIT and MC1R genes and their possible association with the coat color phenotype in this breed, a gene duplication and splice mutation of KIT were diagnosed in a sample of 93 unrelated Rongchang animals. The results show that Rongchang pigs have a single copy of KIT without the splice mutation at the first nucleotide of intron 17, indicating that the dominant white I allele of KIT is not responsible for their white phenotype. The KIT mRNA and MC1R coding sequences were also determined in this breed. Three putative amino acid substitutions were found in the KIT gene between Rongchang and Western white pigs, their association with the Rongchang white phenotype remains unknown. For the MC1R gene, Rongchang pigs were demonstrated to have the same dominant black allele (E(D1)) as other Chinese breeds, supporting the previous conclusion that Chinese and Western pigs have independent domestication origin. We also clarified that the Rongchang white phenotype was recessive to nonwhite color phenotypes. Our results provide a good starting point for the identification of the mutations underlying the white coat color in Rongchang pigs.     

西部地区主要猪种H-FABP基因多态性,IMF含量及不同基因型脂肪细胞脂滴量的关系

H-FABP是FABP家族成员之一,在长链脂肪酸的吸收和代谢平衡中发挥关键作用,但它对猪IMF含量的影响还知之甚少,对中国西部地区的猪种更是如此.文章利用PCR-RFLP（HinfⅠ、HaeⅢ和Msp Ⅰ 3种限制性内切酶）分子标记技术,检测了杜洛克猪、长白猪、大白猪、内江猪、荣昌猪、汉江黑猪、汉中白猪、八眉猪和野猪共计265头猪H-FABP基因5＇-上游区和第二内含子区的遗传变异,利用最小二乘模型分析了H-FABP基因对猪肌内脂肪含量的遗传效应,并运用猪脂肪细胞培养,油红O染色和TG测定等技术检测了H-FABP基因不同基因型脂肪细胞内脂滴的形态和沉积的量.结果表明：（1）在HinfⅠ-RFLP位点上,上述品种和野猪均存在多态性,其中大白猪、八眉猪、汉江黑猪、汉中白猪和野猪表现为低度多态,杜洛克、长白猪、内江猪和荣昌猪为中度多态;除汉江黑猪（P＜0.05）和野猪（P＜0.01）外,其他猪种基因频率和基因型频率都处于Hardy-Weinderg平衡状态（P＞0.05）;而在HaeⅢ-RFLP和Msp Ⅰ-RFLP位点上,仅内江猪、荣昌猪、汉江黑猪和八眉猪为单态;（2）9种基因型对肌内脂肪（IMF）含量的影响,HH＞Hh＞hh,DD＜Dd＜dd,AA＜Aa＜aa,遗传效应值分别为：3.89,3.42,3.17,2.27,2.49,2.91,2.28,2.70,2.95,H-FABP基因可显著地提高IMF含量（P＜0.05）;（3）aaddHH型的脂肪细胞脂滴的形态,密度和含量与其他基因型细胞差异显著（P＜0.05）.结果提示：可通过提高＂aaddHH＂基因型的频率来增加IMF含量,达到改善猪肉质的目的.     

Single nucleotide polymorphism analysis on melanocortin receptor 1 (MC1R) of Chinese native pig

Thecoatcolorisanimportantcharacteristicofapigbreed,andcanbeclassifiedintomanytypes.Thecolorvariationsareeitherduetothedistributionofmelanocytesintheskinortotheabilityofmelano-cytestoproducemelaningranules.Thesynthesisofmelaninoriginatedfromtheformationofneuralcrest-derivedcells,whichhavetwomigrationroutes[1,2].Themelanocytesaretheramificationswhiletheneuralcrest-derivedcellsmigrateviadorsalroute[3].Andtheyprovidethefactoryformelaninsynthesis.Al-thoughtherearemanykindsofpigmentsinvertebralanim…     

Single nucleotide polymorphism analysis on melanocortin receptor 1 (<Emphasis Type="Italic">MC1R</Emphasis>) of Chinese native pig

Melanocortin receptor 1 (MC1R) gene, one of the important candidate genes for coat color trait, was used to analyze the single nucleotide polymorphism (SNP) in Chinese native pig breeds by PCR-single strand conformation polymorphism (PCR-SSCP). The study had also taken 3 imported pig breeds as control. The results showed that the three mutations G284A, T309C and T364C found in Chinese native pigs were consistent to the mutation found in the European Large Black individuals. However, 68CC or C492T and G728A were only found in the imported individuals, which were obviously different from the Chinese native pigs. Accordingly, we presumed that the coat colors of Chinese native pigs belonged to dominant black color system, which was completely distinct to that of imported pig breeds. Thus it was implied thatMC1R gene was not the principal factor affecting the coat color differences of Chinese native pig breeds, but could be used to trace the molecular evolution of pig breeds.     

猪MyoG基因的PCR-RFLP多态性分析

以杜洛克、长白、大约克、南昌白、二花脸、梅山猪、玉山黑猪、乐平花猪、金华两头乌及上高两头乌等中外10个猪种共计561头猪为研究材料,采用3对引物（PCR1、PCR2、PCR3）分别扩增猪肌细胞生成素（MyoG）基因的不同区域,扩增产物经限制性核酸内切酶MspⅠ酶切后发现：（1）在PCR1 MspⅠ-RFLP位点上,外来品种杜洛克、长白、大约克及培育品种南昌白中极大多数个体表现为AA型,个别为BB型;而6个中国地方猪种除乐平花猪外均以BB型居多。（2）在PCR2 MspⅠ-RFLP位点上,6个中国地方猪种除一头玉山黑猪表现为MN型外,其余均为MM型;而外来品种以NN型占大多数,培育品种南昌白更趋向于外来品种。（3）在PCR3 MspⅠ-RFLP位点上,所有猪种均可得到扩增产物,但无MspⅠ酶切位点。（4）在梅山猪及与其亲缘关系较近的二花脸猪中,没有发现Soumillion等(1997)报道的梅山猪特异性MspⅠ多态性酶切位点。     

猪ATF4基因多态性与生产性状的关联及基因表达分析

为进一步了解和认识ATF4基因的功能,揭示ATF4对猪脂肪代谢的影响,寻找与肉质性状相关联的分子标记,文章采用PCR方法扩增了ATF4基因部分序列,通过序列比对发现在翻译起始密码子ATG下游159 bp处存在A159G转换,通过PCR-AluⅠ-RFLP对大白猪、长白猪、梅山猪和通城猪进行酶切分型,发现在大白猪和长白猪中均为AA基因型,在梅山猪和通城猪中均为GG基因型。进一步对大白猪×梅山F2群体资源家系进行了酶切分型,并分析该位点的多态性与生产性状的关系。结果表明,ATF4的多态性与臀部平均膘厚存在极显著相关(P<0.01),与胸腰椎间膘厚、平均膘厚、眼肌高、眼肌面积存在显著相关(P<0.05)。采用Real-time PCR分析了ATF4基因在大白猪与梅山猪背最长肌不同发育阶段的表达模式。结果表明,ATF4基因在大白猪和梅山猪胚胎期65 d和出生后3 d中的表达水平相对都比较低,且在两品种间无明显差异;而在出生后60 d和120 d,ATF4基因在大白猪中与梅山猪均出现了上调表达,并且在梅山猪中的相对表达水平要显著高于大白猪。研究结果为进一步深入研究猪ATF4基因在脂肪代谢中的分子机理奠定了基础。