MC4R、POU1F1基因对京海黄鸡生长性能的遗传效应

以MC4R和POU1F1基因为候选基因, 采用PCR-SSCP和DNA测序技术检测两个候选基因在京海黄鸡群体中的单核苷酸多态性(SNPs), 同时对候选基因与京海黄鸡生长性能的相关性进行了研究。结果表明, MC4R基因编码区第662 bp位置有G→C碱基的点突变, 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0.929, B等位基因频率为0.071; 在POU1F1基因exon3在序列的第5 231 bp位置有一个A→T碱基的点突变, 检测到CC、CD、DD 3种基因型, C等位基因频率为0.500, D等位基因频率为0.500。采用GLM模型分析基因型对生长性能的遗传效应, 结果表明, MC4R基因AA基因型个体的4、8、12周龄体重显著地高于BB型个体(P<0.05), 16周龄体重差异极显著(P<0.01); POU1F1基因CD基因型个体体重极显著高于CC型和DD型(P<0.01)。因此推测MC4R和POU1F1基因可能是影响鸡生长性状的主效基因或与主效基因紧密连锁的标记基因, 能够在分子标记辅助选择中用于对鸡生长性状的遗传改良。     

三个绵羊群体MC4R基因多态性及生物信息学分析

旨在探讨绵羊黑素皮质素受体-4(melanocortin-4 receptor,MC4R)的分子机理,采用PCR-SSCP方法对3个绵羊群体(甘肃肉用绵羊新品种群、小尾寒羊和湖羊)的MC4R基因外显子进行多态性检测和生物信息学分析。结果表明,3个绵羊群体均存在3种基因型AA型、AB型和BB型,优势基因型为BB,其中优势等位基因为B;测序结果表明,野生型BB型和突变型AB型相比,AB型个体在该基因编码区第511位点发生G→A突变,第495位发生C→T突变;AA型个体在该基因编码区第511位点发生G→A突变,出现AA的纯合,第495位发生C→T突变,出现CC纯合;3个绵羊群体中小尾寒羊的多态信息含量属于中度多态(0.25PIC0.50),甘肃肉用绵羊新品种群羊和湖羊属于低度多态(PIC0.25);χ2适合性检验表明除湖羊之外,其余2个绵羊品种均处于Hardy-Weinberg平衡状态。生物信息学分析发现MC4R氨基酸序列有明显的疏水性区域,有7个跨膜螺旋区及信号肽,其编码蛋白主要的二级结构元件是α螺旋和无规则卷曲;同源性比对发现绵羊MC4R基因与山羊、牛、野猪、人类及大猩猩的相似度分别为97%、94%、81%、83%及83%,说明MC4R是一个非常保守的蛋白,在绵羊的生长发育中起着重要作用。     

鸽黑素皮质素受体3、4(MC3R、MC4R)基因多态性与生长性状的相关分析

利用PCR-SSCP技术和DNA测序方法检测广东石岐肉鸽和哈尔滨地区灰色家鸽MC3R和MC4R基因部分编码区序列的单核苷酸多态性, 分析了MC3R基因T91G突变位点和MC4R基因A903G突变位点导致的基因型与两群体鸽生长和体组成性状的关系。结果表明, 这两个多态位点所导致的基因型对石岐肉鸽活重、屠体重、全净膛重均有显著影响(P<0.05); 另外, 利用这两个突变位点所产生的合并基因型在鸽群体中与生长和体组成性状作最小二乘分析, 结果表明, 两位点合并后的基因型对全净膛重影响显著(P<0.05)。多重比较结果表明, BBAA型全净膛重显著大于AABB型, BBAA型对于体重增长是有利基因型。     

四个牦牛品种MC1R基因部分序列的多态性研究

为探索4个牦牛品种MC1R基因多态性的相关信息,选取甘南牦牛、天祝白牦牛、青海高原牦牛、大通牦牛4个品种共408头个体为研究对象,采用PCR-SSCP方法分析牦牛MC1R基因部分序列的基因多态性。结果表明,与GenBank中牛MCIR基因序列(登录号:AF445641.1)比对发现,该扩增片段在3 891 bp处发生C→G的突变,在3 912 bp处发生T→C的突变,共发现CC、DD、EE、CD、CE和DE 6种基因型。4个牦牛品种中CD、CE和DE 3种基因型在青海高原牦牛和大通牦牛中占主要优势,这3种基因型频率总和在青海高原牦牛和大通牦牛群体中分别是0.778和0.781。DD和CD两基因型是甘南牦牛群里中的优势基因型,其基因型频率分别是0.351和0.328。天祝白牦牛中优势基因型是DD,其基因型频率是0.500。D等位基因是4个地方品种牦牛中的优势等位基因。4个地方品种在该基因座上都处于Hardy-Weinberg平衡状态(P>0.05)。青海高原牦牛和大通牦牛两个群体处于高度多态(PIC>0.5),甘南牦牛和天祝白牦牛处于中度多态(0.25相似文献   

犬MC1R基因R306ter与毛色性状相关性研究

目的　分析犬MC1R基因中R30 6ter位点多态性与犬毛色表型的相关性 ,为遗传育种 ,培育出更加优良的实验用犬奠定基础。方法　采用PCR SSCP技术 ,对MC1R基因R30 6ter位点进行基因多态性检测 ,分析位点多态性与毛色性状之间的相关性 ,并对该位点进行克隆测序。结果　PCR SSCP分析结果表明 ,R30 6ter位点序列具有多态性 ,表现为C、D二个等位基因和CC、CD及DD三种基因型。对R30 6ter多态性片段克隆测序发现 ,MC1R基因在编码第 30 6位氨基酸的密码子处存在一个由CGA到TGA的终止突变。结论　经统计分析结果表明在杂种犬中MC1R基因多态性与毛色性状不存在显著的相关性 ,这可能是由于外科手术学教学用犬是杂种犬 ,其遗传背景不同所致。由于MC1R基因的R30 6ter位点内存在碱基变异 ,因此在杂种犬中表现出明显的PCR SSCP多态性     

CXCR2基因多态性与奶牛乳房炎和乳品质的关联

采用PCR-SSCP技术研究了荷斯坦牛、西门塔尔牛和通江黄牛3个品种共160头个体CXCR2基因多态性与乳房炎抗性性状和对牛奶品质性状的遗传效应。结果表明: CXCR2基因有3个SNP多态位点, 分别位于序列的第685 bp、777 bp和861 bp位点, 确定了5个等位基因A、B、C、E和F。685 bp位点表现为BC和CC基因型, 777 bp位点表现为AA和AB基因型, 861 bp位点为CC和BC基因型。与奶牛乳房炎敏感性有关的基因型主要是BC、CC和FF, 可能对乳房炎有抗性的是AA、AB和EE基因型。据不同基因型对乳品质的遗传效应分析来看, AA、AB和EE基因型的乳品质性状极显著或显著优于其他基因型。     

猪TLR4基因外显子1新等位基因的分离及遗传变异分析

文章采用PCR-SSCP方法对亚洲野猪、3个引进的商业化品种和10个中国地方猪品种共893个个体TLR4基因外显子1的遗传变异进行了检测,旨在系统分析国内外猪种TLR4基因的多态性,为探讨该基因在免疫和防御系统中发挥的作用提供依据。结果,在猪TLR4基因外显子1中分离到新的等位基因,共检测到3个等位基因,6种基因型。其中杜洛克检测到AA、BB、CC、AB、AC、BC基因型,有杜洛克血统的苏太猪中检测到BB、CC、BC基因型,长白猪、约克夏中检测到CC、BC基因型,野猪及所有10个中国地方猪品种TLR4基因外显子1高度保守,只检测到CC基因型,中国地方猪品种和引进品种TLR4基因外显子1多态性存在极显著的差异。3种基因型中CC型与GenBank中的序列一致,BB和AA基因型分别存在G93C同义突变位点和G194A无义突变位点,这2个变异位点与抗逆性和一般抗病力的关系值得进一步深入研究。     

藏獒MC1R基因多态性与毛色表型的关联研究

目的:通过检测藏獒黑素皮质激素受体1(MC1R)基因的单链构象多态性(SSCP)在不同毛色群体中的分布,探讨MC1R基因多态性与毛色表型的相关性。方法:采用DNA测序技术,选择不同毛色藏獒的DNA为样本,根据GenBank发布的荷斯坦牛MC1R基因序列设计一对引物,采用PCR-SSCP技术分析MC1R基因在藏獒中的SSCP。结果:MC1R基因在藏獒中具有PCR-SSCP多态性,分别检测到3种基因型(AA、AB和BB);对MC1R基因多态性片段DNA克隆测序后发现,MC1R基因在编码区第313位存在单碱基突变(G→A),该突变导致第105位氨基酸发生由丙氨酸向苏氨酸的改变(T105A)。结论:MC1R基因的多态性与毛色性状不存在显著的相关性。     

GHR基因第10外显子与塞北兔生长发育性能相关性研究

本研究利用单链构象多态性(SSCP)和测序技术,对塞北兔群体GHR基因第10外显子进行多态性研究,并与生长发育性能进行关联性分析。结果发现:有两处点突变,214处为同义突变,均编码苯丙氨酸;AB基因型在255处出现A位点的缺失,引起移码突变;塞北兔整个群体呈现哈代-温伯格平衡状态,但母兔群体不平衡;AA基因型比AB基因型对塞北兔活体重,胴体重,前腿重均有提高,差异显著(p0.05)。表明:母兔群体在育种过程中受到人工选择而引起遗传漂变,AA基因型为有利基因型,因此可作为影响塞北兔生长性状的主效候选基因,进行早期辅助选择。     

麦洼牦牛和九龙牦牛FSHβ基因的PCR-SSCP分析

运用一对特异性引物对麦洼牦牛、九龙牦牛的FSHβ基因的5’端侧翼区进行了扩增,应用PCR-SSCP方法对其进行了多态性分析。结果表明:在麦洼牦牛的FSHβ基因的5’端侧翼区有AA型、AB型和BB型三种基因型,九龙牦牛的FSHβ基因的5’端侧翼区只检测到了AA型、AB型两种基因型。在两种牦牛品系中,AA基因型频率最高,A等位基因频率均明显高于B等位基因频率。麦洼牦牛和九龙牦牛群体中多态信息含量分别为0.3431、0.1411,麦洼牦牛多态性能高,遗传变异大。     

Polymorphism of bone morphogenetic protein 4 gene and its relationship with litter size of Jining Grey goats

Two pairs of primers (P1 and P2) were designed to detect single nucleotide polymorphisms of exon 2 and intron 2 of bone morphogenetic protein 4 (BMP4) gene in both high fecundity breed (Jining Grey goat) and low fecundity breeds (Boer, Angora and Inner Mongolia Cashmere goats) by single strand conformation polymorphism. Results showed that no polymorphism was detected for exon 2 (primer P1) of BMP4 gene in four goat breeds. For intron 2 (primer P2), three genotypes (AA, AB and BB) were detected in Jining Grey and Inner Mongolia Cashmere goats, two genotypes (AB and BB) in Angora goats, and only one genotype (AA) in Boer goats. Sequencing revealed one mutation (2203G>A) of BMP4 gene in the genotype BB in comparison to the genotype AA. The differences of litter size between AA, AB and BB genotypes were not significant (P > 0.05) in Jining Grey goats. A pair of primer (P3) was designed to detect polymorphism in the 3' flanking region of BMP4 gene that contained dinucleotide repeated sequence (CA) in the four goat breeds by microsatellite analysis. For primer P3, three genotypes (CC, CD and DD) were detected in four goat breeds. Sequencing revealed one more CA dinucleotide in genotype DD than in genotype CC. The Jining Grey does with genotype CC had 0.55 (P < 0.05) or 0.72 (P < 0.05) kids more than those with genotype CD or DD. These results preliminarily indicated that allele C of BMP4 gene is a potential DNA marker for improving litter size in goats.     

Association between PCR-SSCP of bone morphogenetic protein 15 gene and prolificacy in Jining Grey goats

The Jining Grey is a prolific local goat breed in P.R. China. Bone morphogenetic protein 15 (BMP15) gene that controls high fecundity of Inverdale, Hanna, Lacaune, Belclare, Cambridge, and Small Tailed Han ewes was studied as a candidate gene for the prolificacy of Jining Grey goats. According to the sequence of ovine BMP15 gene, six pairs of primers were designed to detect single nucleotide polymorphisms in exon 1 and exon 2 of the BMP15 gene in both high fecundity breed (Jining Grey goats) and low fecundity breeds (Boer, Liaoning Cashmere, and Inner Mongolia Cashmere goats) by single strand conformation polymorphism (SSCP). Two pairs of primers (F1/R1 and F2/R2) were used to amplify the exon 1. Four pairs of primers (F3/R3, F4/R4, F5/R5, and F6/R6) were used to amplify the exon 2. Only the products amplified by primer F5/R5 displayed polymorphism. Results indicated that two genotypes (AA and AB) were detected in prolific Jining Grey goats and only one genotype (AA) was detected in low fecundity goat breeds. In Jining Grey goats frequencies of genotypes AA and AB were 0.10 and 0.90, respectively. Sequencing revealed two point mutations (G963A and G1050C) of BMP15 gene in the AB genotype in comparison to the AA genotype. In Jining Grey goats the heterozygous AB does had 1.13 (p < 0.01) kids more than the homozygous AA does. These results preliminarily showed that the BMP15 gene is either a major gene that influences the prolificacy of Jining Grey goats or a molecular genetic marker in close linkage with such a gene.     

GDF9 as a candidate gene for prolificacy of Small Tail Han sheep

Growth differentiation factor 9 (GDF9) which controls the fecundity of Belclare, Cambridge, Santa Ines, Moghani, Ghezel and Thoka ewes was studied as a candidate gene for the prolificacy of Small Tail Han sheep. According to the sequence of ovine GDF9 gene, six pairs of primers were designed to detect single nucleotide polymorphisms of two exons of GDF9 gene in both high fecundity breed (Small Tail Han sheep) and low fecundity breed (Dorset sheep) by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). Only the products amplified by primers 2-1 and 2-2 displayed polymorphisms. For primer 2-1, three genotypes (AA, AB and BB) were detected in both sheep breeds. Sequencing revealed one silent mutation (G477A) in exon 2 of GDF9 gene in the BB genotype in comparison with the AA, which was known as G3 mutation of GDF9 gene in Belclare and Cambridge ewes. The relationship of least squares means for litter size was AA?>?AB?>?BB in Small Tail Han sheep (P?>?0.05). For primer 2-2, two genotypes (CC and CD) were detected in both sheep breeds. Sequencing revealed one novel single nucleotide mutation (G729T) in exon 2 of GDF9 gene in the CD genotype in comparison with the CC, which resulted in an amino acid change (Gln243His). The ewes with mutation heterozygous genotype CD had 0.77 (P?相似文献   

The association of two single nucleotide polymorphisms (SNPs) in growth hormone (GH) gene with litter size and superovulation response in goat-breeds

Two active mutations (A 781 G and A 1575 G) in growth hormone (GH) gene, and their associations with litter size (LS), were investigated in both a high prolificacy (Matou, n = 182) and a low prolificacy breed (Boer, n = 352) by using the PCR-RFLP method. Superovulation experiments were designed in 57 dams, in order to evaluate the effect of different genotypes of the GH gene on superovulation response. Two genotypes (AA and AB, CC and CD) in each mutation were detected in these two goat breeds. Neither BB nor DD homozygous genotypes were observed. The genotypic frequencies of AB and CC were significantly higher than those of AA and CD. In the third parity, Matou dams with AB or CC genotypes had significantly larger litter sizes than those with AA and CD (p < 0.05). On combining the two loci, both Matou and Boer dams with ABCD genotype had the largest litter sizes when compared to the other genotypes (p < 0.05). When undergoing like superovulation treatments, a significantly higher number of corpora lutea and ova, with a lower incidence of ovarian cysts, were harvested in the AB and CC genotypes than in AA and CD. These results show that the two loci of GH gene are highly associated with abundant prolificacy and superovulation response in goat breeds.     

FSHβ 基因PCR-SSCP多态性及其与济宁青山羊高繁殖力关系的研究

采用PCR-SSCP技术检测促卵泡素b亚基(follicle-stimulating hormone β, FSHβ)基因5′调控区、外显子1和外显子2在高繁殖力山羊品种(济宁青山羊)和低繁殖力山羊品种(辽宁绒山羊、波尔山羊、安哥拉山羊)中的单核苷酸多态性, 同时研究该基因对济宁青山羊高繁殖力的影响。结果表明: 山羊与绵羊的FSHβ 基因该段核苷酸序列同源性为98%。9对引物中, 只有P9的扩增片段存在多态性。P9的扩增片段在济宁青山羊和辽宁绒山羊中检测到AA、AB和AC 3种基因型; 在波尔山羊中检测到AA、CC和AC 3种基因型; 在安哥拉山羊中检测到AA、BB、CC、AB、AC和BC共6种基因型。测序分析发现BB型与AA型相比在外显子2的第94 bp处有G→A突变, 并引起氨基酸改变(丙氨酸→苏氨酸); CC型与AA型相比在外显子2的第174 bp有一处C→T沉默突变。济宁青山羊AA、AB和AC基因型频率分别为0.686、0.137和0.177。AA基因型济宁青山羊产羔数最小二乘均值比AB基因型的多0.78只(P<0.05), 比AC基因型的多0.64只(P<0.05)。     

Polymorphisms of BMPR-IB gene and their relationship with litter size in goats

The bone morphogenetic protein receptor IB (BMPR-IB) gene was studied as a candidate gene for the prolificacy of goats. According to mRNA sequence of ovine BMPR-IB gene, ten pairs of primers were designed to detect single nucleotide polymorphisms (SNPs) of exon 1, exon 2, exon 6 to exon 10 and 3′ untranslated region (UTR) of the BMPR-IB gene in both high prolificacy breed (Jining Grey goat) and low prolificacy breeds (Wendeng Dairy and Inner Mongolia Cashmere goats) by polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) method. Only the products amplified by primers P8 and P10 of the 3′UTR displayed polymorphisms. For primer P8, three genotypes (AA, AB and BB) were detected in Jining Grey and Wendeng Dairy goats, two genotypes (AA and AB) were in Inner Mongolia Cashmere goats. Sequencing revealed one mutation (71C→T) of the BMPR-IB gene in genotype BB compared with AA. The differences of least squares mean (LSM) for litter size between genotypes AA, AB and BB were non-significant (P > 0.05) in Jining Grey goats. For primer P10, three genotypes (CC, CD and DD) were detected in Jining Grey and Wendeng Dairy goats and one genotype (CC) in Inner Mongolia Cashmere goats. Sequencing revealed one mutation (130T→C) of the BMPR-IB gene in genotype DD compared with CC. The differences of LSM for litter size between genotypes CC, CD and DD were non-significant (P > 0.05) in Jining Grey goats. These results preliminarily showed that the detected loci of the BMPR-IB gene had no significant effect on prolificacy of Jining Grey goats.     

A 15 nucleotide deletion mutation in coding region of the RIG-I lowers grass carp (Ctenopharyngodon idella) resistance to grass carp reovirus

RIG-I (Retinoic acid-inducible gene I) is a pivotal receptor that detects numerous RNA and DNA viruses and plays crucial roles in the induction of type I interferons. In the present study, a deletion mutation in CiRIG-I (Ctenopharyngodon idella RIG-I) coding region was detected, its association with resistance/susceptibility to grass carp reovirus (GCRV) was examined, and possible mechanism was analyzed. A 15-bp deletion mutation was found, and the mutation results in a deletion of five amino acids. To investigate the genotypes and alleles, the relevant PCR products were electrophoresed on 2.5% agarose gel. Three genotypes and two alleles were discovered. The general allele was named as A and the deletion mutation allele was named as B. The deletion mutation cancels a predicted phosphorylation site and changes the secondary structure and the probability of peroxisomal targeting signal 1 in CiRIG-I. To explore the correlation between these genotypes and the resistance of grass carp to GCRV, a challenge experiment was carried out. The cumulative mortality in genotype AA (40.70%) and AB (52.73%) was significantly lower than that in genotype BB (71.43%) (P = 0.032). The result demonstrated that genotype AA and AB were resistant to GCRV, while genotype BB was susceptible. The 15-bp deletion mutation lowers the resistance of grass carp to GCRV. This result might provide a potential genetic marker for further investigation of selective breeding of resistant grass carp to GCRV.     

FGF5基因对内蒙古绒山羊绒毛性状的影响

根据FGF5基因已知DNA序列设计了2对引物, 采用PCR-SSCP和PCR-RFLP技术, 在内蒙古绒山羊群体中进行基因多态性检测, 结果发现FGF5基因外显子1存在限制性内切酶BglⅠ多态位点。对其不同基因型个体PCR回收产物进行测序, 测序结果发现该SNP是由碱基序列C→T的突变而引起的。基因型和基因频率统计, 该实验群体以等位基因A具有明显的优势, χ2检验表明该SNP位点的基因频率处于Hardy-Weinberg平衡状态; 对该SNP与绒毛性状关联分析, 表明该SNP对绒纤维伸直长度(P<0.01)和含绒量(P<0.05)有显著影响, 而对其他各绒毛性状的影响不显著(P>0.05)。AB基因型个体绒纤维伸直长度(P<0.01)和含绒量(P<0.05)显著高于AA基因型个体。     

The CSN1S1?N and F alleles identified by PCR-SSCP and their associations with milk yield and composition in Chinese dairy goats

A method was depicted to identify null allele CSN1S1 N and low allele CSN1S1 F of the CSN1S1 gene of goat using PCR-SSCP (polymerase chain reaction-single strand conformation polymorphism). First, primer A was designed to amplify the exon 9 of CSN1S1 gene which produced three genotypes AA, AB, and BB. Among these three genotypes, only AA and AB individuals had a cytosine deletion at exon 9 after DNA sequencing, which cannot be used to identify the N and F alleles. Therefore, primer B was used to amplify intron 14 of CSN1S1 of described AA and AB individuals. Genotypes FF, FN and NN were detected within AA individuals and genotypes FO and NO were detected in the above AB individuals. The frequencies of F and N alleles in 708 samples from Xinong Saanen (XS) and Guanzhong (GZ) dairy goat breeds were 0.1139, 0.0927, and 0.2376, 0.1193, respectively. In 268 XS samples, the individuals with NN genotype contained a significant lower protein content than that of other genotypes (P<0.01). Individuals of FF genotype had significant higher milk yield than that of NO genotype in the first milk lactation of 202 XS individuals (P<0.05). Therefore, the variability at CSN1S1 locus contains enough genetic diversity to be potentially useful in improving the quality and production of milk in Chinese dairy goat breeds.