Study on the Relationship between the Polymorphisms of PRLR Gene and Lactation Traits in Jiangyue Donkeys

In this study, the single nucleotide polymorphism of prolactin receptor (PRLR) gene and its correlation with the lactation traits in Jiangyue donkeys was tested, and the molecular genetic marker of breeding Jiangyue donkeys was explored.120 individuals were analyzed association of PRLR gene polymorphism with lactation traits in Jiangyue donkeys. Genetic polymorphism of PRLR gene flanking region was detected by PCR-SSCP method. The results showed that there was 1 mutation in PRLR gene and 2 genotypes of AA and AB, and allele A was the dominant allele. The SNP g.29764584 C>G was identified of PRLR gene by sequencing, and it was a synonymous mutation, did not cause amino acid changes. Statistical results indicated that the SNP was significantly associated with average daily milk yield and protein percentage in Jiangyue donkey (P<0.05),there was no significant in the lactin percentage(P>0.05). The mutation might be as crucial DNA genetic marker for lactation performance selection in Jiangyue donkeys.     

白羽番鸭脂联素基因SNP与肌内脂肪和血清总胆固醇的关联分析

采用PCR-SSCP法对白羽番鸭脂联素基因全编码区和内含子进行多态性检测,并分析其多态对肌内脂肪(IMF)和血清总胆固醇(TC)的遗传效应,为番鸭分子标记辅助选择提供参考.结果表明:3个SNP分别在外显予1和外显子2的编码区发生A167G和G711A突变,为沉默突变,在内含子发生C290T突变;3个SNP位点均处于Ha...     

疆岳驴催乳素受体基因多态性及其与泌乳性状间的相关性分析

试验旨在研究疆岳驴催乳素受体（prolactin receptor,PRLR）基因多态性及其与泌乳性状间的关系,探寻可用于疆岳驴选育的分子遗传标记。随机选取120头疆岳驴,对疆岳驴的泌乳性能指标进行测定,并运用PCR-SSCP方法对疆岳驴PRLR基因侧翼区进行多态性分析。结果显示,PRLR基因在疆岳驴群体中存在2种基因型：AA和AB,A为优势等位基因;测序结果显示,PRLR基因的碱基突变位置为g.29764584 C>G,该突变为同义突变,没有导致编码的氨基酸发生改变;经关联分析,PRLR基因AB基因型个体平均日泌乳量显著高于AA基因型（P<0.05）,AA基因型个体乳蛋白率显著高于AB基因型（P<0.05）,在乳糖率上差异不显著（P>0.05）。疆岳驴PRLR基因突变与泌乳性状存在关联性,推测PRLR基因可以作为疆岳驴乳用型选育的分子遗传标记之一。     

玉树马和德保矮马SLC36A1基因多态性研究

 控制马香槟毛色的CH（Champagne gene）基因家族包含4个候选基因（SLC36A1、SLC36A2、SLC36A3、SPARC）,研究发现SLC36A1基因外显子2的突变是造成马香槟毛色的关键位点。为揭示中国马SLC36A1基因遗传多态性,本研究以玉树马和德保矮马共74个样本为研究对象,以马DNA池为模板扩增SLC36A1基因的10个外显子及部分内含子序列并进行测序分析。共发现马SLC36A1基因5个SNPs,分别位于内含子3（g.26699953 A>G, g.26699851 G>C, g.26699850 G>C）,外显子4（g.26699562 G>A）及外显子6（g.26697018 C>T）。利用PCR RFLP方法对74个家马样本进行基因分型,发现外显子6的SNP有基因型CC、CT;外显子4的SNP有基因型GG、GA;内含子3的g.26699850 G>C突变有基因型GG、GC;内含子3的另外2个SNPs（g.26699953 A>G, g.26699851 G>C）通过测序,发现有AA、AG、GG与GG、GC、CC基因型。所有5个马SNPs均为野生型占主要优势。由此界定了马SLC36A1基因有9种单倍型（H1 H9）,其中H5是最主要的单倍型。德保矮马遗传多样度为0.4190,比玉树马（0.2228）高,表明德保矮马香槟毛色遗传多态性比玉树马更丰富。玉树马与德保矮马的平均单倍型多样度为0.3160,表明其香槟毛色遗传多态性相对较低。     

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.     

Association of a novel SNP in exon 10 of the IGF2 gene with growth traits in Egyptian water buffalo (Bubalus bubalis)

Insulin-like growth factor 2 (IGF2) plays an important role in muscle growth and it might be used as a marker for the growth traits selection strategies in farm animals. The objectives of this study were to detect polymorphisms in exon 10 of IGF2 and to determine associations between these polymorphisms and growth traits in Egyptian water buffalo. PCR-single-strand conformation polymorphism (SSCP) and DNA sequencing methods were used to detect any prospective polymorphism. A novel single nucleotide polymorphism (SNP), C287A, was detected. It was a non-synonymous mutation and led to replacement of glutamine (Q) amino acid (aa) by histidine (H) aa. Three different SSCP patterns were observed: AA, AC, and CC, with frequencies of 0.540, 0.325, and 0.135, respectively. Association analyses revealed that the AA individuals had a higher average daily gain (ADG) than other individuals (CC and AC) from birth to 9 months of age. We conclude that the AA genotype in C287A SNP in the exon 10 of the IGF2 gene is associated with the ADG during the age from birth to 9 months and could be used as a potential genetic marker for selection of growth traits in Egyptian buffalo.     

京海黄鸡IGFBP-3基因外显子1和内含子1部分序列多态性及其与生长繁殖性状的相关性

以京海黄鸡母鸡为材料,采用PCR-SSCP技术对IGFBP-3基因外显子1及内含子1部分序列多态位点进行研究,并计算基因型频率、基因频率、卡方值和部分遗传多态性指标。结果表明,在外显子1上没有检测到多态位点,在内含子1上检测到1个多态位点,该位点为中度多态,在160 bp处发生T到G的突变,导致BB、AB型的56日龄体重显著大于AA型(P＜0.05),AA型的11月产蛋数显著大于BB、AB型(P＜0.05)。由此初步推断,内含子1对产蛋及生长发育很可能有一定的促进作用,A为产蛋的优势基因,B为体重的优势基因。     

4个绵羊品种Leptin基因部分片段的SNPs分析

利用PCR-SSCP技术对萨福克、道塞特、特克塞尔及滩羊4个绵羊品种358个个体Leptin基因2、3外显子进行多态性分析,共检测到7个SNPs,其中新发现5个SNPs。测序结果表明,在外显子2上无突变。内含子2上存在99位碱基由A突变为G;115位碱基由G突变为A;150位碱基由C突变为T;171位碱基由C突变为T。外显子3上,存在271位碱基由G突变为A,使编码的Arg转变为Gln;316位碱基由C突变为A,使编码的Pro转变为Gln;387位碱基由G突变为T,使编码的Val转变为Leu。     

绵羊瘦素受体基因部分序列测定及其变异位点分析

试验旨在检测绵羊瘦素受体(leptin receptor,LEPR)基因序列突变位点,为进一步分析LEPR基因多态性与绵羊生长性状的关系奠定基础。选用美利奴羊与阿华西羊杂交群体作为试验材料,利用RT-PCR、测序等方法测定了4只绵羊的LEPR基因cDNA部分序列及内含子7序列,同时利用PCR-RFLP分析两个位点在群体(229只)中的多态性。测定出LEPR基因cDNA序列长2608 bp(包含外显子2-16完整序列及外显子1和17的部分序列)和LEPR第7内含子序列全长160 bp,共发现5个核苷酸变异位点,第2外显子中2个(T240C和A279G),第10、14外显子中各1个(A1683G,T2373C),内含子7中1个(1285+A73G),外显子中的4个变异位点均未引起编码氨基酸的改变。在研究群体中,A279G与A1683G中A的频率分别为0.415、0.467,后者处于非平衡状态,GG和AA是主要的单倍型。不同物种间序列一致性分值均比较高,但LEPR mRNA区序列一致性比内含子高,基于LEPR mRNA序列构建的系统发育树更符合实际情况,且可靠性值更高。结果表明,绵羊LEPR基因的保守性较强,突变形式主要是转换,A279G和A1683G可能是突变热点。     

兴义鸭肌肉生长抑制素基因多态性与屠宰性状的相关性分析

为了解兴义鸭肌肉生长抑制素(myostatin,MSTN)基因SNPs与屠宰性状的相关性,本研究采用基因克隆及PCR产物直接测序的方法,将MSTN基因作为鸭屠宰性状的候选基因,对兴义鸭的MSTN基因外显子进行多态性检测.结果表明,在60个兴义鸭个体中筛选出8个SNPs,其中,第1外显子有5个突变位点:SNP1(G77A)、SNP2(A91G)、SNP3(G130A)、SNP4(C325T)和SNP5(C331T);在第2外显子中并未发现突变位点;第3外显子有3个突变位点:SNP6(C206T)、SNP7(A235G)和SNP8(C256A);对这8个SNPs与屠宰性状进行关联性分析,结果并未达到显著水平(P>0.05).本研究结果可丰富MSTN基因的研究数据,为鸭的育种提供参考.     

Novel Variants in the HMGA2 Gene Are Associated With Withers Height in Debao Pony

The Debao pony is a well-known dwarf horse breed in China. High-mobility group AT-hook 2 (HMGA2) gene is regarded as one of the important candidate genes regulating body height in horses. The aim of this study was to study the association between mutations in HMGA2 gene and withers height in Debao ponies. The polymorphisms in all exons and partial introns of the HMGA2 gene were screened with sequencing across 180 Debao ponies. And the association between the DNA variants and withers height was analyzed. Seven genetic variants were identified in HMGA2 gene, including six novel variants. Among them, six mutations were located in two closed linked blocks. The three novel variants (In1-1, E5-1, and E5-2) in the 1st intron and the fifth exon and a known mutation (In1-2) had significant association with withers height in Debao ponies. These results suggest that the four variants have the potential to be used as genetic markers for dwarf horse breeding activities.     

4个绵羊品种Leptin基因部分片段的SNPs多态性与生长发育性状的相关性分析

利用PCR-SSCP技术对萨福克、陶赛特、得克塞尔及滩羊4个绵羊品种358个个体Leptin基因等2、3外显子进行多态性分析,共检测到7个SNPs,其中新发现5个SNPs。测序结果表明,在外显子2上无突变。内含子2上存在A99G、G115A、C150T、C171T位点。外显子3上,存在G271A;C316A;G387T位点。外显子3上的SNPs使编码的氨基酸发生变化。统计分析表明A99G、C150T和A99G＋C150T位点与生长发育性状存在相关性。在A99G位点,Aa基因型初生质量、日增质量、体高、胸围和尻宽指标上均高于AA基因型,初生质量、日增质量和体高指标差异显著（P〈0．05）,胸围和尻宽指标差异极显著（P〈0．01）。C150T和A99G＋C150T位点结果一致,突变基因型日增重、体高、体长、胸围和尻宽指标均高于野生基因型,差异显著（P〈0．05）。     

哈萨克羊DRB1基因外显子3多态性及生物信息学分析

试验旨在研究白细胞表面抗原DRB1基因外显子3多态性与哈萨克羊布鲁氏菌病易感性的相关性。运用混合DNA池结合PCR产物直接测序方法,对哈萨克羊DRB1基因外显子3进行多态性分析,经卡方检验分析每个SNP位点的等位基因频率、基因型频率及其多态性与布鲁氏菌病易感性的相关性,利用生物信息学分析软件对PCR扩增所获序列进行RNA二级结构及蛋白质的二级结构和抗原表位分析。结果表明,在282 bp的外显子3序列中共检测到7个SNPs,分别为：T10C、C119T（Trp→Arg）、G215C（Gln→Glu）、A238G、T245G（Ser→Ala）、G256A、C259T,这些位点在病例组和对照组之间的等位基因频率及各基因型间不存在显著性差异（P > 0.05）;进一步分析发现,各突变位点均引起RNA二级结构和最小自由能的改变,各错义突变位点均未引起蛋白质二级结构和抗原表位的改变。由此得出,DRB1基因外显子3的7个SNPs位点（T10C、C119T、G215C、A238G、T245G、G256A和C259T）与哈萨克羊布鲁氏菌病易感性无相关性。     

Polymorphisms and Its Genetic Variation Analysis of DQB2 Gene Exon 2 in Kazakh Sheep

This study was aimed to investigate the association between the polymorphism of DQB2 gene exon 2 and the susceptibility to Kazakh sheep brucellosis. The DQB2 gene exon 2 of Kazakh sheep lymphocyte antigen was amplified by PCR-SSCP method from 146 healthy and 28 infected with Brucella Kazakh sheep, and the single nucleotide polymorphisms (SNP) was analyzed, then the different alleles were selected for cloning and sequencing. In order to analyze its correlation with brucellosis susceptibility, the differences in gene frequency and genotype frequency of each SNP locus were analyzed by Chi-square test. Bioinformatics softwares were used to analyze the secondary structure of mRNA, the secondary structure, tertiary structure and epitope of protein. The sequencing result showed that 33 SNPs were detected in 270 bp DNA sequence, the gene frequencies of C9G and A180G were extremely significantly different in case group and control group (P<0.01), and its genotype frequencies presented significantly difference (P<0.05). Similarly, A13T and C133G loci were significant difference in case group and control group (P<0.05). Further analysis result showed that the minimum free energy of the A180G mutation site was the lowest and its mRNA secondary structure was the most stable; Both A13T and C133G mutation sites caused the changes of mRNA secondary structure, protein secondary, tertiary structure and antigenic epitope of protein, respectively. The results showed that the polymorphism of DQB2 gene exon 2 might be significantly correlated with brucellosis susceptibility in Kazakh sheep.     

哈萨克羊DQB2基因外显子2多态性及遗传变异分析

试验旨在研究DQB2基因外显子2多态性与哈萨克羊布鲁氏菌病易感性的相关性。通过PCR-SSCP技术对146只布鲁氏菌阴性哈萨克羊血液样本和28只布鲁氏菌阳性哈萨克羊血液样本中的白细胞表面抗原DQB2基因外显子2的多态性进行研究,挑取不同的等位基因进行克隆测序,经卡方检验分析每个SNP位点的基因频率、基因型频率及其多态性与布鲁氏菌病易感性的相关性,应用生物信息学软件分析与哈萨克羊布鲁氏菌病易感性相关的不同等位基因的mRNA二级结构及蛋白质的二级结构、三级结构和抗原表位。结果发现,在270 bp的外显子2序列中共检测到33个SNPs,其中C9G、A180G位点的基因频率在病例组和对照组中的分布具有极显著性差异（P<0.01）,其基因型频率在病例组和对照组中存在显著差异（P<0.05）;A13T、C133G位点的基因频率在病例组和对照组中存在显著差异（P<0.05）;进一步分析发现,A180G突变位点的最小自由能最低,其mRNA二级结构最稳定;A13T和C133G 2个突变位点均引起mRNA二级结构及蛋白质二级结构、三级结构和抗原表位的改变。本试验结果表明DQB2基因外显子2多态性与哈萨克羊布鲁氏菌病易感性呈显著相关。     

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。     

贵州半细毛羊ADAMTS1基因部分外显子多态性研究

为探索ADAMTS1基因在绵羊繁殖中的作用,本研究以贵州半细毛羊为试验对象构建DNA池,分别设计引物扩增第1、2、5-7和9外显子,研究ADAMTS1基因的遗传多态性.结果显示,ADAMTS1基因第6外显子有1个SNP (A255C),第9外显子有4个SNPs (A127G、G243T、G303T、A401G).生物信息学分析表明,SNPs位点对ADAMTS1基因RNA二级结构和蛋白结构均有一定影响.ADAMTS1基因可能影响贵州半细毛羊的繁殖性状.     

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.