不同牛种STAT5B基因启动子多态性分析

目的:试验旨在筛选STAT5B基因启动子区SNP及研究其对启动子功能元件的影响,为地方牛种选种选育提供一定理论依据.方法:选择品种差异较大的贵州荷斯坦奶牛和务川黑牛构建不同DNA池,直接测序筛选SNP位点.结果:STAT5B基因5'调控区及第1外显子存在3个SNPs位点,分别为:T-181C、C-31G、T+119C.生物信息学软件预测得到STAT5B基因核心启动子区和转录因子结合位点,SNP位点导致5个转录因子结合位点消失,而产生8个新的转录因子结合位点.软件未发现可能的CpG岛范围,但STAT5B基因RNA二级结构和最小自由能在突变后显著改变.结论:DNA池结合测序技术可快速筛选SNP位点,STAT5B启动子区存在功能性SNP位点.     

中国四品种黄牛性染色体多态性的研究

对4个地方黄牛品种（蒙古牛,秦川牛,岭南牛和西镇牛）共63头牛（37♂,26♀）染色体核型的比较研究表明,性染色体在品种间和品种内均存在多态性。Y染色体有中、亚中和近端着丝点染色体,其频率有一定的分布规律。X染色体均为一大的亚中着丝点染色体,但臂比和着丝点指数有品种差异。本文从性染色体多态性上揭示了中国黄牛起源的多元性,为中国黄牛品种的系统分类增添了又一新的依据和线索。     

Y-SNPs Haplotype Diversity in Four Chinese Cattle Breeds

To investigate the genetic diversity of Chinese cattle, 96 male samples of 4 Chinese native cattle breeds were investigated using 5 single nucleotide polymorphisms specific to the bovine Y chromosome. Two previously described haplotypes (taurine Y2 and indicine Y3) were detected in 74 and 22 animals, respectively. The haplotype frequencies varied amongst the four native breeds. The taurine Y2 haplotype dominated in the Qinchuan, Dabieshan, and Yunba breeds. However, the indicine Y3 haplotype occurred in high frequency in the Enshi breed. Among the four native breeds, Yunba had the highest haplotype diversity (0.4330 ± 0.0750), followed by Qinchuan (0.2899 ± 0.1028) and Enshi (0.2222 ± 0.1662), Dabieshan was the least differentiated (0.1079 ± 0.0680). Compared with some foreign cattle breeds, the low level of haplotype diversity was detected in our breeds (0.2633 ± 0.1030).     

10个牛品种线粒体12S rRNA基因多态性分析

以鲁西牛、渤海黑牛、大别山牛、南阳牛、晋南牛、秦川牛、中国西门塔尔牛、日本和牛、海福特牛和安格斯牛10个牛品种共计353头牛为研究对象,利用单链构象多态性（SSCP）分子标记方法对其线粒体DNA 12S rRNA基因进行了多态性分析,结果发现该基因出现分布频率不同的3种单倍型（Ⅰ、Ⅱ、Ⅲ）,其中单倍型Ⅰ出现的频率最低,仅在晋南牛1个个体中发现;单倍型Ⅱ在中国本地黄牛中都存在,在中国西门塔尔牛中的分布频率很低,但在3个国外品种海福特牛、安格斯牛和日本和牛中不存在;单倍型Ⅲ在这10个牛品种中均有分布,但以国外品种最丰富。计算平均多态信息含量,发现我国地方品种在0.232～0.423之间,基本属于中度多态,说明我国地方黄牛品种在12S rRNA上多态性比较丰富,国外品种较为贫乏;但是单倍型Ⅰ频率很低有丢失的趋势,需要对其进行保护。     

Polymorphism of the Goat Agouti Signaling Protein Gene and Its Relationship with Coat Color in Italian and Spanish Breeds

Agouti signaling protein (ASIP) is one of the key players in the modulation of hair pigmentation in mammals. Binding to the melanocortin 1 receptor, ASIP induces the synthesis of phaeomelanin, associated with reddish brown, red, tan, and yellow coats. We have sequenced 2.8?kb of the goat ASIP gene in 48 individuals and identified two missense (Cys126Gly and Val128Gly) and two intronic polymorphisms. In silico analysis revealed that the Cys126Gly substitution may cause a structural change by disrupting a highly conserved disulfide bond. We studied its segregation in 12 Spanish and Italian goat breeds (N?=?360) with different pigmentation patterns and found striking differences in the frequency of the putative loss-of-function Gly(126) allele (Italian 0.43, Spanish Peninsular 0.08), but we did not observe a clear association with coat color. This suggests that the frequency of this putative loss-of-function allele has evolved under the influence of demographic rather than selection factors in goats from these two geographical areas.     

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   

三个山羊品种KAP8基因的PCR-SSCP分析

目的：为了筛选绒山羊产绒性状的候选基因。方法：采用PCR-SSCP法对三个山羊品种角蛋白辅助蛋白8（KAP8）进行了多态性研究。结果：三个品种中KAP8.1均未出现多态,而KAP8.2均出现了三种带型：AA、BB和AB。Hardy-Weinberg分析表明辽宁绒山羊和黎城大青羊未达到平衡状态,辽-苛高代杂种山羊达到了平衡。测序结果显示,和Genebank登陆序列相比,AA和BB均发生了碱基变化。其中AA型出现了以下变异：216bp（G-A）,217（A-G）,BB型出现了以下变异：216bp（G-A）,232bp（C-T）。分析表明AA型变异造成了氨基酸序列发生了变化（R-K）,而BB型突变未造成氨基酸发生改变。结论：KAP8.2可能是影响绒山羊产绒性状的基因之一。     

Genetic Polymorphism of β-Lactoglobulin Gene in Native Turkish Sheep Breeds

The genetic polymorphism of the β-lactoglobulin gene was investigated in three native Turkish sheep breeds. The study was carried out on 108 sheep (29 Kıvırcık, 38 G?k?eada, and 41 Sakız) by means of PCR-RFLP methods. Two genetic variants (A and B) and three genotypes (AA, AB, and BB) of β-lactoglobulin have been identified. The gene frequencies of β-LG A and B were 0.7759 and 0.2241 in Kıvırcık, 0.7632 and 0.2368 in G?k?eada, and 0.9756 and 0.0244 in Sakız breeds, respectively. The populations were in Hardy–Weinberg equilibrium in all samples from the three breeds.     

山东地方山羊BMP15基因多态性与产羔数性状关联分析

利用PCR、克隆测序、序列拼接获得山羊(Capra hircus)BMP15基因全长。利用F-CSGE技术分析两个外显子,发现山羊BMP15编码序列的第901处发生了A→G单碱基突变,该突变使得第301位氨基酸(成熟蛋白质第32位氨基酸)由丝氨酸变为甘氨酸。利用LDR技术对济宁青山羊、鲁北白山羊和沂蒙黑山羊进行突变检测,并进行其与产羔数的关联分析。结果表明,该突变对济宁青山羊产羔数没有显著影响,但对鲁北白山羊及沂蒙黑山羊产羔数均有显著影响(P0.05)。GG型和AG型的鲁北白山羊产羔数分别比AA型多0.34只(P0.01)和0.31只(P0.01)。AG型沂蒙黑山羊的产羔数比AA型多0.13只(P0.01)。初步表明,BMP15是控制鲁北白山羊和沂蒙黑山羊多胎性状的一个主效基因或是与之存在紧密遗传连锁的分子标记。     

RAPD Analysis of DNA Polymorphism in Turkish Sheep Breeds

The genetic diversity and phylogenetic relationships of 108 individual sheep from three Turkish sheep breeds (K?v?rc?k, Gökçeada, and Sak?z) were studied using RAPD analysis. Polymorphisms within and between populations were assayed using 15 random primers, and 82 loci were amplified ranging from 250 to 2,500 bp. The percentage of polymorphic loci was found to be 80.49, 78.05, and 73.17%, for K?v?rc?k, Gökçeada, and Sak?z sheep breeds, respectively. Total genetic diversity was 0.2265, and the average coefficient of genetic differentiation was 0.1181. Genetically, the Gökçeada breed was more closely related to the Sak?z breed than to the K?v?rc?k breed.     

不同猪种E.coli F18受体基因的多态性

采用PCR—RFLP技术检测了大约克、长白、杜洛克、宁乡、沙子岭和大围子6个品种共867头猪的E.coli F18受体(ECF18R)基因座的遗传变异。结果表明：Hin6 Ⅰ-RFLP位点上,大约克、长白、杜洛克3个外来猪种均存在多态,且以敏感型(GG型和AG型)居多,平均占94％,3个外来猪种的G等位基因频率平均为0．76,AA抗性型个体占少数,平均为6％,猪群中M307处G→A的突变频率并不高。宁乡、沙子岭和大围子3个本地猪种的所有检测样品都表现为GG型,在该位点上均不存在G→A的突变。各猪种ECF18受体基因座的PCR—RFLP基因型分布X^2检验结果表明,每个外来猪种ECF18受体基因座的PCR—RFLP基因型分布与3个本地猪种的相比均差异显著或极显著,3个本地猪种间的ECF18受体基因座的PCR—RFLP基因型分布完全一致。外来猪种间只有长白与杜洛克各基因型的分布差异显著,其余均不显著。     

5个山羊品种CSN1S2基因的Alw26I酶切

利用PCR-RFLP技术对西农萨能奶山羊、关中奶山羊、陕南白山羊、安哥拉山羊和波尔山羊5个山羊品种的170个个体的αs 2酪蛋白(CSN1S2)基因进行多态性分析,结果表明：扩增大小为310 bp的片段经限制性内切酶Alw26I酶切后表现多态,且5个山羊品种该基因座位均处于Hardy-Weinberg平衡状态。西农萨能奶山羊、关中奶山羊、陕南白山羊、安哥拉山羊和波尔山羊的基因杂合度/有效等位基因数/Shaanon信息熵/PIC值分别为0.1589/1.1889/0.2955/0.1463, 0.4114/1.6981/0.6017/0.5171,0.1653/1.1980/0.3046/0.1516,0.0646/1.0691/0.1463/0.0625,0.0541/1.0572/ 0.1270/ 0.0526。分析结果显示,关中奶山羊的遗传多样性最丰富,表现为高度多态;其次是西农萨能奶山羊和陕南白山羊,而安哥拉山羊和波尔山羊的遗传变异程度最低。     

Evaluation of Genetic Differentiation in Bos indicus Cattle Breeds from Marathwada Region of India Using Microsatellite Polymorphism

Elucidation of genetic variability and genetic relationship among breeds has direct relevance with the issues of sustainable use of domestic animal genetic resources. In the present study, genetic polymorphism was evaluated using 22 microsatellite loci in unrelated samples of Red Kandhari and Deoni cattle breeds inhabiting the same geographical area of Marathwada region in Maharashtra state (western India). This work was mainly aimed at assessing the current genetic diversity to understand whether the two zebu populations in question are genetically differentiated. A total of 164 alleles were detected with an average of 5.82 and 5.86 alleles per locus (MNA) in Red Kandhari and Deoni breeds, respectively. The estimated mean observed (Ho) and expected (He) heterozygosity were 0.47 and 0.64 in Red Kandhari vs. 0.57 and 0.69 in Deoni cattle, respectively, demonstrating considerable level of genetic variation in both the populations. Mean estimates of F statistics were: F (FIT) = 0.315±0.035, f(FIS) = 0.231±0.031, θ(FST) = 0.110±0.022, with both the breeds exhibiting significant deficit of heterozygotes (FIS = 0.179 in Deoni; 0.278 in Red Kandhari). The multilocus FST values implied that 11.0% of the total genetic variation corresponds to breed and were statistically greater than zero for the two populations, suggesting population division. The evaluation of exact test also indicated that allele frequencies across all the loci differed significantly (P < 0.001) between two zebu breeds, further supporting population differentiation. Different genetic distance measures showed considerable levels of distances between the two cattle breeds (0.318 = Nei's standard DS; 0.250 = Nei's DA; 0.416 = Cavalli-Sforza and Edwards's DC; 0.164 = Reynold's, and 2.64 = Delta mu square (dμ)2. Bayesian statistical approach to assign each individual to the population also supported considerable differentiation between the two cattle breeds, possibly reflecting the limited gene flow between the two Marthwada cattle populations. The existence of cohesive breeding structure of both the breeds was further substantiated by allele-sharing distance measures (DAS) among individual animals. The results of this study thus revealed that the two Bos indicus breeds sharing the common breeding tracts are genetically differentiated enough as separate breeds.     

奶牛TLR4基因的多态性与乳房炎的相关性研究

TLR4通过识别病原体而激活免疫细胞,在先天免疫和适应性免疫防御中起着重要作用。以中国荷斯坦奶牛、三河牛和中国西门塔尔牛为研究对象,以TLR4为乳房炎抗性的候选基因,分别扩增316bp和382bp2个片段,分别采用SSCP和RFLP-AluⅠ方法来检测TLR4基因的多态性。结合测序发现:在intron1的第4,525bp处的A→G的突变,和exon3的第1,397bp处的T→C突变,使得产生多态。2个位点的A、B等位基因在3个群体中都有分布,且处于中度多态。χ2适合性检验表明,3个群体在这2个位点的突变达到Hardy-Weinberg平衡状态(P>0.05)。运用SAS8.0软件采用最小二乘法拟合线性模型,将基因座不同基因型与奶牛乳房炎进行了关联分析,结果表明:T4CRBR1的AA基因型为乳房炎抗性基因型(P<0.05),A等位基因为乳房炎抗性的有利基因,T4CRBR2的各基因型个体间的体细胞评分差异不显著(P>0.05)。     

3个鸡种GARS-AIRS-GART基因第21号外显子的SNPs检测

采用PCR-SSCP和DNA测序法对安卡鸡、文昌鸡和如皋鸡中GARS-AIRS-GART基因的第21号外显子进行SNPs检测.结果发现该外显子含4个核苷酸多态位点:G29943A、C29968T、T30011C和C30017T,其中3处为非同义突变,分别为:29 943处天冬氨酸(Asp)→天冬酰胺(Asn)、30 011处色氨酸(Trp)→精氨酸(Arg)、30 017处精氨酸(Arg)→半胱氨酸(Cys);另一处为同义突变.对多态位点进行单倍型分析,发现12种单倍型,其中两种单倍型(5号和6号)的频率超过10%,分别为0.330 0和0.162 8,1号、3号、9号和10号单倍型频率介于1%～10%之间,其余都小于1%.另外共检测到11种基因型,存在5种等位基因,安卡鸡的D和E等位基因的基因频率较高,基因型与其它两个鸡品种之间都存在着极显著的差异(P<0.01),表明我国地方鸡种和外来鸡种在遗传组成上存在着差异.本研究将为鸡肉品质相关研究积累基础数据,为今后深入开展肉品质的开发利用奠定理论基础.     

Genetic Polymorphism of Type 1 Intermediate Filament Wool Keratin Gene in Native Indian Sheep Breeds

Information is presented on the genetic polymorphism of the Type 1 intermediate filament wool keratin gene in 15 native Indian sheep breeds belonging to different agro-ecological regions of India. The study analyzed random blood samples of the 638 sheep by the PCR-RFLP technique. Restriction digestion analysis of a 480 bp PCR fragment of the first exon region with MspI revealed two allelic variants (M = 0.748 and N = 0.252) and three genotypes (MM = 0.543, MN = 0.410, and NN = 0.047) across the 15 sheep breeds. The allelic frequency differences for both alleles across the Indian breeds, irrespective of their geographic distribution, color pattern, and utility traits, were observed to be statistically insignificant by a chi-square test (P > 0.05). According to the pattern of occurrence of allelic variants (M > N), the Indian breeds exhibited similarity to some of the reported European sheep breeds. The average heterozygosity was 0.420, and none of the breeds deviated from Hardy-Weinberg equilibrium. The predominance of the M over the N allele supported its ancestry in Indian sheep too.