郏县红牛IGF2基因多态性与生长性状关系研究

本研究利用PCR-SSCP技术首次对郏县红牛母牛IGF2基因多态性及其与生长性状关系进行了研究。结果表明:IGF2基因第2外显子表现多态性,存在AA、AB、BB三种不同基因型。其频率分别为0.235/0.480/0.285;A/B等位基因频率分别为0.474/0.526;该位点纯合度为0.501,杂合度为0.499,位点间有效等位基因数为1.996。该基因位点处于Hardy-Weinberg平衡状态(P<0.05)。通过对郏县红牛IGF2基因第2外显子位点不同基因型与样本体重及其它体尺指标的关联分析表明:不同基因型对胸围(P<0.01)和体重(P<0.05)有显著影响,AB基因型个体的胸围和体重明显大于AA和BB型个体。说明IGF2基因可以作为标记用于郏县红牛的肉用选育。     

STAT5b基因2个SNPs位点与京海黄鸡生长和繁殖性状的关联分析

采用PCR-SSCP方法检测京海黄鸡、AA鸡、尤溪麻鸡、边鸡4个鸡品种STAT5b基因5′调控区部分序列的多态性,并分析其对京海黄鸡生长和繁殖性状的遗传效应,利用PHASE软件对2个多态位点进行单倍型分析。在5′调控区检测到C-1591T、G-250A 2个SNPs;单倍型分析产生4种单倍型H1(CG)、H2(TG)、H3(CA)、H4(TA),单倍型组合H1H4对初生重有极显著影响(P0.01);单倍型组合H3H4与8、16周龄体重存在正相关(P0.05),单倍型组合H1H2对开产蛋重有极显著影响(P0.01);单倍型组合H3H4的开产体重极显著高于其他单倍型组合(P0.01);在300日龄体重上,单倍型组合H1H1和H1H3显著高于H4H4(P0.05)。京海黄鸡STAT5b基因5′调控区的多态性对其生长和繁殖性状有一定影响,但是否可以作为影响京海黄鸡生产性状的遗传标记需进一步研究。     

鸡胰岛素样生长因子1受体基因G26336A、C111014A多态位点的检测及其与生产性状的相关性

根据GenBank发表的鸡胰岛素样生长因子1受体(insulin-like growth factor 1 receptor,IGF1R)基因序列针对外显子2和3设计2对引物,采用DNA池测序和PCR-SSCP技术,对鸡IGF1R基因的G26336A、C111014A位点进行多态性检测,并与鸡的生产性状进行相关性分析。结果表明,G26336A位点AA基因型个体56日龄体重显著低于BB基因型个体(P<0.05),AA和AB基因型个体在84日龄体重显著低于BB基因型个体(P<0.05)。C111014A位点CC、CD基因型个体初生重显著低于DD基因型(P<0.05),CC基因型个体在300日龄体重显著低于DD基因型(P<0.05)。研究结果初步表明,IGF1R基因可能是影响鸡生长性状的一个主效基因或是与之存在紧密遗传连锁的一个标记。     

京海黄鸡IGF1R基因Alu Ⅰ、Hin1 Ⅰ多态位点的发现及其与生产性状的相关性

根据GenBank发表的鸡IGF1R基因序列针对外显子2和3设计2对引物,采用池DNA测序和PCR-R FLP技术,对京海黄鸡IGF1R基因的AluⅠ、Hin1Ⅰ位点进行多态性检测,并与京海黄鸡的生产性状进行相关性分析。结果表明:AluⅠ位点AA基因型个体在8周龄体重显著低于BB型个体(P<0.05),AA和AB基因型个体在12周龄体重显著低于BB型个体(P<0.05)。Hin1Ⅰ位点CC、CD基因型个体的初生重显著低于DD基因型(P<0.05),CC基因型个体的300日龄体重显著低于DD基因型(P<0.05)。研究结果初步显示,IGF1R基因可能是影响京海黄鸡生长性状的一个主效基因或是与之存在紧密遗传连锁的一个标记。     

MSTN基因外显子3的多态性及其与边鸡生长性状的关联分析

采用PCR-SSCP技术检测边鸡及2个对照鸡群体(京海黄鸡和尤溪麻鸡)肌肉生长抑制素(myostatin, MSTN)基因外显子3的单核苷酸多态性,并与边鸡的生长性状进行相关性分析。结果表明,在MSTN基因外显子3的8527—8528 bp处检测到一个TC的插入。在边鸡和京海黄鸡中只检测到2种基因型(AA和AB基因型),在尤溪麻鸡中检测到3种基因型(AA、AB和BB基因型)。在3个鸡品种中都以A等位基因为优势等位基因。最小二乘分析结果表明,边鸡2种基因型个体的6～16周龄体重均无显著差异(P>0.05)。     

Novel polymorphisms of the IGF1R gene and their association with average daily gain in Egyptian buffalo (Bubalus bubalis)

The objective of this study was to detect insulin-like growth factor 1 receptor (IGF1R) polymorphisms, their allele, and genotype frequencies and to determine associations between these polymorphisms and growth traits in Egyptian water buffalo. Three loci of the IGF1R coding region were amplified by RT-PCR and, subsequently, subjected to sequence analysis, followed by single-strand conformation polymorphism to identify different allelic patterns. A total of 11 novel polymorphisms were detected; 6 SNPs among Egyptian water buffaloes and 5 polymorphisms compared with Indian buffalo (Y12700). Three of those polymorphisms; GAG Indel polymorphism, C261G, and G263C SNPs, were nonsynonymous mutations. The GAG Indel polymorphism led to deletion of E (glutamic) amino acid (aa) in the IGF1R of Egyptian water buffaloes compared with Indian buffalo. However, C261G SNP, which replaced A (alanine) by G (glycine) aa, and G263C SNP, which changed A (alanine) to P (proline) aa, were detected among Egyptian water buffaloes. Three different single-strand conformation polymorphism patterns were observed in exon 21: CC/CC, GG/GG, and CG/GC with frequencies of 0.291, 0.253, and 0.556, respectively. The heterozygous animals (CG/GC) had a higher ADG than homozygous animals (CC/CC and GG/GG) from birth to 6 mo of age. We conclude that the heterozygous haplotype, C261G/G263C, in exon 21 of the IGF1R gene is associated with the ADG during the early stages of life (from birth to 6 mo of age) and could be used as a genetic marker for selection of growth traits in Egyptian buffalo.     

Identification of a single nucleotide polymorphism of the insulin-like growth factor binding protein 2 gene and its association with growth and body composition traits in the chicken

Insulin-like growth factor binding protein 2 regulates a broad spectrum of biological activities involved in growth, development, and differentiation. The current study was designed to investigate the associations of IGFBP2 gene polymorphisms with chicken growth and body composition traits. The Northeast Agricultural University Resource Population (NEAURP) was established by crossing broiler sires, derived from lines at Northeast Agricultural University, that were divergently selected for abdominal fat, with Baier layer dams, a Chinese local breed. The F1 birds were inter-crossed to produce an F2 population. Body weight and body composition traits were measured in the NEAURP. The PCR primers for the intron 2 region of IGFBP2 were designed based on chicken genomic sequence. Nucleotide polymorphisms between parental lines were detected by DNA sequencing. A C/T SNP in intron 2 was detected, and PCR-RFLP methods were then developed to genotype the F2 individuals. The results showed that the IGFBP2 SNP was associated with multiple traits, including BW, metatarsus length, shank length, femur length, shank weight, femur weight, metatarsus claw weight, and abdominal fat weight in the 1,028 NEAURP F2 individuals. This research suggests that IGFBP2 or a tightly linked gene has broad effects on growth and development in the chicken.     

Study on association of single nucleotide polymorphism of CAPN1 gene with muscle fibre and carcass traits in quality chicken populations

This study was aimed at investigating the effect of the calpain 1 (CAPN1) gene on carcass and meat quality traits in eight meat-type chicken populations, including five pure lines (developed from Chinese local breeds) and three cross-breeds. Primer pairs for the Coding Sequence (CDS) region in CAPN1 were designed from the chicken genomic sequence database. Polymorphisms were detected by polymerase chain reaction (PCR)-Single Strand Conformation Polymorphism (SSCP) and DNA sequencing. Three single nucleotide polymorphisms (SNP; C2546T, G3535A and C7198A) were detected among individuals in each population. The associations of their haplotypes (H1 = CGA, H2 = CGC, H3 = CAA, H4 = CAC, H5 = TGA and H7 = TAA) with chicken breast muscle fibre and carcass traits were analysed. Results showed that the haplotypes were associated with live weight (LW), carcass weight (CW), breast muscle weight (BMW) and leg muscle weight (LMW) (p < 0.05), and were also related to eviscerated percentage (%EP) and breast muscle fibre density (p < 0.01). H1H3 haplotype was dominant for LW, CW and BMW; H1H5 haplotype was dominant for EP; H3H4 haplotype was dominant for LMW and H1H1 haplotype was dominant for BFD. It was concluded that the CAPN1 gene may be a major gene affecting meat quality traits of chicken or it is linked with the major gene. H1H3, H1H5 and H3H4 were the most advantageous haplotypes for carcass traits whereas H1H1 was the positive haplotype for breast muscle fibre trait.     

Correlation Analysis Between Haplotypes of ApoA5 Gene on Body Weight and Meat Quality in Jinfen White Pigs

This study was aimed to explore the correlation between the haplotypes of apolipoprotein A5 (ApoA5) gene and body weight,meat quality in Jinfen White pigs.180 Jinfen White pigs were selected,PCR was used to detect the single nucleotide polymorphism (SNP) of ApoA5 gene,and the linkage of these SNPs was analyzed by Haploview software.Non-fully linked SNPs were screened to form the haplotypes of ApoA5 gene,and the frequency of different haplotypes of ApoA5 gene were calculated.The correlation between the haplotypes of ApoA5 gene and body weight,meat quality was analyzed by SPSS software to identify the haplotypes of ApoA5 gene with excellent traits.The results showed that ApoA5 gene of Jinfen White pigs contained 21 SNPs and 5 completely linked regions which included 5,2,3,4 and 2 SNPs,respectively.Ten non-completely linked tag SNPs were selected to form the haplotypes of ApoA5 gene,Hap 1 had the highest gene frequency (46.7%).The haplotypes of ApoA5 gene had a significant correlation with the birth weight,weaning weight,6-month-old body weight,intramuscular fat content,shear force and drip loss,but no significant correlation with pH_{24 h} in Jinfen White pigs.Hap 11 of ApoA5 gene in Jinfen White pigs had the highest birth weight,weaning weight,6-month-old body weight and intramuscular fat content,the lowest shear force,and the lower drip loss.Hap 6 had the lowest birth weight.Hap 2 had the lowest weaning and 6-month-old weight.Hap 3 had the lowest intramuscular fat content and the highest shear force.Hap 10 had the highest drip loss.Hap 11 of ApoA5 gene in Jinfen White pigs had excellent traits,which could be used as a molecular marker to use in molecular breeding of Jinfen White pigs and improve the breeding efficiency.     

晋汾白猪ApoA5基因单倍型与体重和肉质性状的相关性分析

试验旨在探究晋汾白猪载脂蛋白A5（apolipoprotein A5,ApoA5）基因的单倍型与体重和肉质性状的相关性。选取180头晋汾白猪为研究群体,利用PCR技术检测ApoA5基因的单核苷酸多态性（single nucleotide polymorphism,SNP）,并采用Haploview软件分析这些SNP位点的连锁不平衡程度;筛选非完全连锁的SNP位点,构建不同的ApoA5基因单倍型,统计不同单倍型的基因频率,并通过SPSS软件分析不同ApoA5基因单倍型与晋汾白猪体重和肉质性状的相关性,鉴定优良性状的ApoA5基因单倍型。结果显示,晋汾白猪ApoA5基因共检测到21个SNPs位点,其中5个完全连锁单倍型块,分别包含5、2、3、4和2个SNPs位点;另外10个非完全连锁的标签SNP组合成不同的ApoA5基因单倍型,其中Hap 1的基因频率最高,达到46.7%,远高于其他单倍型;ApoA5基因单倍型与晋汾白猪的初生重、断奶重、6月龄体重、肌内脂肪含量、剪切力和滴水损失均具有明显的相关性,而与pH_{24 h}没有明显的相关性;ApoA5基因Hap 11晋汾白猪的初生重、断奶重、6月龄体重和肌内脂肪含量均最高,且剪切力最低,滴水损失也相对较低;Hap 6的初生重最低;Hap 2的断奶重和6月龄体重均最低;Hap 3的肌内脂肪含量最低,剪切力最高;Hap 10的滴水损失最高。晋汾白猪ApoA5基因Hap 11属于优良性状单倍型,是一个具有优良性状的分子标记,可以用于晋汾白猪的分子育种,提高选种效率。     

Fine-mapping of quantitative trait loci for body weight and bone traits and positional cloning of the RB1 gene in chicken

Previously, a quantitative trait locus (QTL) that affects body weight (BW) at 4-12 weeks of age and carcass weight at 12 weeks of age had been mapped on chicken chromosome 1. After including more markers and individuals, the confidence interval was narrowed down to approximately 5.5 Mbps and located this QTL near a microsatellite marker (ADL328). This QTL is the same as the QTL for 12 bone traits, including metatarsus length and metatarsus circumference at 4, 6, 8, 10 and 12 weeks of age and keel length and metatarsus claw weight at 12 weeks of age, that was identified using the same population. In the current study, 1010 individuals from the Northeast Agricultural University F(2) resource population were used and 14 single-nucleotide polymorphism (SNPs) around ADL328 were developed to construct haplotypes, and an association analysis was performed to fine-map the QTL. The haplotypes were constructed on the basis of a sliding 'window', with three SNP markers included in each 'window'. The association analysis results indicated that the haplotypes in 'windows' 6-12 were significantly associated with BW and bone traits and suggested that the QTL for BW and bone traits was located between SNP8 and SNP14 or was in linkage disequilibrium with this region. The interval from SNP8 to SNP14 was approximately 400 kbps. This region contained five RefSeq genes (RB1, P2RY5, FNDC3A, MLNR and CAB39L) on the University of California Santa Cruz website. The RB1 gene was selected as a candidate gene and five SNPs were identified in the gene. The association results indicated that the RB1 gene was a major gene for BW and bone traits. The SNPs g.39692 G>A and g.77260 A>G in RB1 gene might be two quantitative trait nucleotides for BW and bone traits.     

The SNP genotypes of growth hormone gene associated with reproductive traits in Tsaiya ducks

Our previous cDNA microarray study showed that the growth hormone (GH) gene may involve in the duck egg formation process. The purpose of this study was to investigate the relationship between GH genotypes of single nucleotide polymorphisms (SNPs) and reproductive traits of Tsaiya ducks. Primer pairs for the coding region in the GH were designed based on the duck genomic sequence. Polymorphisms were detected by polymerase chain reaction (PCR)-single strand polymorphism (SSCP) and were verified by DNA sequencing. Nineteen SNPs were identified in the duck GH gene, of which three coding SNPs (C3169T, C3700T and C5058G) were genotyped to investigate the associations with reproductive traits. The results showed that each SNP was associated with at least one duck fertility-related trait (p < 0.05). Haplotypes constructed on these three SNPs were associated with fertility rate (FR) and maximum duration of fertility (MDF) (p < 0.05). In particular, diplotype H1H1 was dominant for FR and MDF. This suggested that GH gene polymorphisms are associated with duck fertility-related traits. The SNPs in this gene may be used as potential markers for marker-assisted selection.     

中国美利奴羊DLX3基因3′UTR的多态性及其与羊毛品质性状的关联分析

为探讨DLX3基因的多态性与绵羊羊毛品质及生长性状的关系,本研究以5个品系的中国美利奴羊(新疆军垦型)为试验材料,采用PCR-RFLP方法开展了DLX3基因3′UTR区4个SNPs的多态性检测。通过卡方检验分析了4个SNPs在各品系绵羊中的等位基因频率,采用连锁不平衡分析构建了这4个SNPs的单倍型,最后将单位点和单倍型分别与绵羊羊毛品质和生长性状进行关联分析。结果表明:4个SNPs在5个品系间的等位基因频率均存在极显著差异(P<0.01);超细毛绵羊品系与其他4个品系间的基因型分布均存在极显著差异(P<0.01),2个多胎品系与其他3个品系间的基因型分布同样存在极显著差异(P<0.01);相关分析显示,这4个SNPs及其单倍型均对羊毛卷曲度有显著影响(P<0.05),而对其他羊毛性状及生长性状无显著影响(P>0.05)。由此可见,中国美利奴羊(新疆军垦型)DLX3基因3′UTR区的多态性与绵羊毛发卷曲度性状显著相关,可以尝试使用这些SNPs开展高品质细毛羊的分子标记辅助选择。     

IGF-I与IGFBP-1基因对京海黄鸡生长性状的遗传效应分析

旨在对IGF-I和IGFBP-1基因部分SNPs与鸡生长性状进行关联分析。试验以京海黄鸡为材料,采用PCR-SSCP方法检测2个候选基因的单核苷酸多态性(SNPs),采用一般线性模型(GLM)分析基因型与生长性状的遗传效应。结果表明,IGF-I基因外显子3序列的60bp处有A→G的点突变,在京海黄鸡中检测到AA、AB、BB 3种基因型,A等位基因频率为0.613,B等位基因的频率为0.387;IGFBP-1基因外显子2序列的21bp处有A→T的点突变,104bp处有T→C的突变,在京海黄鸡中检测到CC、CD和DD 3种基因型,C等位基因频率为0.558,D等位基因频率为0.442。IGF-I基因BB基因型个体4周龄体质量显著高于AA和AB型个体(P<0.05);IGFBP-1基因DD基因型个体8周龄体质量显著高于CC基因型个体(P<0.05),4、12和16周龄体质量差异极显著(P<0.01)。因此,推测这些SNPs对京海黄鸡生长性状具有一定的影响,应用于鸡育种过程中的标记辅助选择可以加快育种进程。     

贵州小香鸡GH基因SNP与生长性状的相关性分析

采用PCR直接测序法对贵州小香鸡GH基因的2～3外显子区域进行SNPs检测,测定部分生长性状并分析其与基因多态性间的关联性。结果显示,在内含子2中共检测出2个变异位点,均存在3种基因型。最小二乘分析显示C170T多态位点显著影响贵州小香鸡的屠宰率(P<0.05),初生重、1周龄重、2周龄重和龙骨长存在显著的基因型与性别互作效应(P<0.05);T334C多态位点与贵州小香鸡体斜长、胫长和胫围存在显著的基因型与性别互作效应(P<0.05)。结果表明,C170T和T334C多态位点可能对贵州小香鸡生长性状有显著影响。     

边鸡IGF-ⅠR基因AluⅠ位点多态性及其对生长性状和繁殖性状的遗传效应分析

为了寻找与边鸡生长性状和繁殖性状相关的遗传标记,本试验采用PCR-RFLP技术检测边鸡类胰岛素生长因子Ⅰ受体（insulin-like growth factor Ⅰreceptor,IGF-ⅠR）基因Alu Ⅰ位点的多态性,并分析该多态位点对边鸡生长性状和繁殖性状的遗传效应。结果显示,边鸡IGF-ⅠR基因Alu Ⅰ位点存在多态性,在外显子2的376 bp处有1个G→A突变,产生了GG、GA和AA 3种基因型,基因型频率分别为0.687、0.296和0.017,GG为优势基因型;G等位基因频率为0.835,为优势等位基因。卡方适合性检验结果表明,该基因座位在边鸡群体中处于哈代-温伯格平衡状态（P>0.05）。相关分析结果表明,在8、14、16和18周龄时,GG基因型个体的体重显著高于GA基因型（P<0.05）,GG基因型个体的开产体重极显著高于GA基因型个体（P<0.01）,但并不能说明该位点可以作为影响边鸡繁殖性状的遗传标记。IGF-ⅠR基因外显子2的Alu Ⅰ位点（G→A）可能是影响边鸡生长性状的一个遗传标记。     

Study on the Polymorphisms of IGF-ⅠR Gene Alu Ⅰ Locus and Genetic Effect on Growth and Reproduction Traits in Bian Chicken

Single nucleotide polymorphisms (SNPs) in exon 2 of insulin-like growth factor Ⅰ receptor (IGF-ⅠR) gene were detected by PCR-RFLP in Bian chicken,which was to find the correlation markers of growth and reproduction traits in Bian chicken.The results showed that AluⅠsite displayed polymorphic, and a G→A mutation at the position of the 376 bp in exon 2 was occurred. There were three genotypes (GG, GA and AA), the genotypes frequencies were 0.687, 0.296 and 0.017, respectively, GG genotype was the dominant genotype, the frequency of G allele was 0.835, which was the dominant allele. This site was in Hardy-Weinberg equilibrium (P>0.05).Correlation analysis showed that GG genotype had significantly higher body weight than those of GA genotype at the age of 8, 14, 16 and 18 weeks (P<0.05);The GG geno type had extremely significantly higher body weight at first egg than those of GA genotype (P<0.01),but it didn't mean that this locus could be used as a genetic marker for reproduction traits in Bian chicken. This results suggested that the AluⅠsite (G→A) in exon 2 of IGF-ⅠR gene could be used as a genetic marker for growth traits in Bian chicken.     

促卵泡激素受体基因第2内含子多态性对如皋黄鸡繁殖性状的遗传效应分析

试验以260只如皋黄鸡为素材,采用飞行质谱技术对位于促卵泡素受体(follicle stimulate hormone receptor,FSHR)基因第2内含子的3个SNPs位点进行检测,并对其与繁殖性状的相关性进行了分析。结果表明,C+29329A突变对开产日龄、开产蛋重及40周龄蛋重有显著影响(P<0.05);而C+30582G只与40周龄总产蛋数呈显著相关(P<0.05);G+51411A则对开产日龄、开产体重及40周龄总产蛋数有显著影响(P<0.05)。研究结果显示,FSHR基因有望成为选育高产蛋鸡的分子标记,辅助选育、开发中国地方鸡品种资源。     

槟榔江水牛ACTA1基因多态性与生长性状的相关性研究

[目的]揭示α-肌动蛋白1（actin alpha 1,ACTA1）基因多态性与槟榔江水牛生长性状的相关性.[方法]采用PCR扩增及基因测序技术研究了槟榔江水牛ACTA1基因多态性,并分析了该基因变异与水牛生长性状的关联关系.[结果]检测到槟榔江水牛ACTA1基因存在2个SNP位点,分别位于第5内含子与第6外显子,水牛ACTA1基因变异与体重、管围等性状存在显著相关.[结论]本研究首次检测了槟榔江水牛ACTA1基因多态性与生长发育性状的关联关系,为槟榔江水牛遗传资源保护利用及标记辅助选择提供了理论依据.     

Genetic Effects of IGF-Ⅰ Gene on the Growth and Reproductive Traits in Jinghai Yellow Chicken

The aim of this study was to analyze the association of SNPs in insulin like factor-Ⅰ（IGF-Ⅰ） gene with chicken growth and reproductive traits. PCR-SSCP approach was applied to assess the single nucleotide polymorphisms(SNPs) and the general linear model was used to analyze genetic effects between genotypes and growth traits of the Jinghai Yellow chicken. For the IGF-Ⅰ gene, three genotypes(AA, AB and BB) were detected in Jinghai Yellow chicken population, the frequency of A and B alleles were 0.613 and 0.387. Sequencing revealed one mutation(A60G) of IGF-Ⅰ gene in BB genotype in comparison to AA genotype. The results showed that BB genotype of the IGF-Ⅰ gene had higher bodyweight at 4 weeks compared to AA and AB genotypes(P<0.05);AA genotype of the IGF-Ⅰ gene had significantly higher bodyweight at 300 day-age-weight compared to BB genotype(P<0.05). Polymorphisms in exon 3 of IGF-Ⅰ gene had certain effect on growth and reproductive traits in Jinghai Yellow chicken. However, whether these polymorphisms were important genetic markers, which related with IGF-Ⅰ gene and reproduction traits in Jinghai Yellow chicken, were needed to be further studied.