共查询到20条相似文献，搜索用时 31 毫秒
1.
不同品种马ELA-DQA* exon2的多态性分析 总被引：5，自引：0，他引：5
通过PCR—SSCP技术检测了7个品种300匹马ELA—DQA＊^exon2的多态性。用12％非变性聚丙烯酰胺凝胶电泳将已变性的PCR扩增产物分离，银染法显色。结果300匹马中共出现27种基因型：10种纯合子和17种杂合子；7个品种马的He和PIC值均表现为高度多态；等位基因序列分析得到10个等位基因间的Kimura双参数遗传距离为0．015～O．147，符合（dN：〉dS）的正选择检验。因此表明马的ELA—DQA＊^exon2多态性丰富。 相似文献
2.
为了解普氏野马、哈萨克马与焉耆马的ELA-DRA*exon2的多态性以及等位基因频率分布。本研究采用PCR-SSCP技术对普氏野马、哈萨克马与焉耆马的ELA-DRA*exon2进行分型,并对不同等位基因进行核苷酸与氨基酸分析。结果显示:127匹马中共出现7种基因型,3种纯合子分别记为AA、BB、CC;4种杂合子分别记为AB、AC、BC、AD。AA基因型为哈萨克马与焉耆马的优势基因型,普氏野马以CC基因型为主。经χ2检验后,3种类型马的等位基因频率和基因型频率均处于Hardy-Weinberg平衡状态。PIC值与He值分析表明,3种类型马均属于中度多态,且普氏野马的PIC值与He值比哈萨克马与焉耆马低。 相似文献
3.
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。 相似文献
4.
WANG Yuan-yuan YAN Guo LUO Cheng QI Jiang-jiao ZHOU Guang-pu TAN Jun GAO Jian-feng 《中国畜牧兽医》2017,44(12):3543-3553
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. 相似文献
5.
本试验采用PCR-SSCP方法对148只布鲁氏菌阴性和60只布鲁氏菌阳性中国美利奴羊白细胞表面抗原DQB1(OLA-DQB1)基因exon 2单核苷酸多态性(SNPs)进行了检测,之后挑选不同等位基因进行PCR产物测序,旨在确定该基因的多态性位点,并对每个SNP位点的等位基因频率、基因型频率进行统计分析,从而分析其多态性与布鲁氏菌病易感性的相关性.测序结果表明,在270 bp的序列内共检测到43个SNPs,其中G196A位点的等位基因频率在病例组和对照组中的分布存在极显著差异(P< 0.01),其基因型频率存在显著差异(P< 0.05);C211T位点的等位基因频率在病例组和对照组中存在显著差异(P< 0.05).由此表明,OLA-DQB1基因exon 2多态性与中国美利奴羊布鲁氏菌病易感性呈显著相关. 相似文献
6.
WANG Wen-wen XU Wan-yun HU Meng-wei LI Jian-hua LIU Peng-tao GAO Jian-feng 《中国畜牧兽医》2015,42(6):1495-1503
The single nucleotide polymorphisms (SNPs) of ovine lymphocyte antigen DQB1 (OLA-DQB1) gene exon 2 was amplified by PCR-SSCP method from 148 healthy and 60 infected with Brucella Chinese Merino sheep and then PCR products of different alleles were sequenced to determine the polymorphism loci of the gene.The differences in gene frequency and genotype frequency of each SNP loci were analyzed statistically to analyze its correlation with brucellosis susceptibility.The sequencing result showed that 43 SNPs were detected in 270 bp DNA sequence,the gene frequencies of G196A allele had extremely significant difference in case and control samples (P< 0.01),and its genotype frequencies presented significant difference (P< 0.05).Similarly,C211T allele was significantly different in case and control samples (P< 0.05).The results showed that the polymorphism of OLA-DQB1 gene exon 2 might be a significant association gene with brucellosis susceptibility. 相似文献
7.
为分析肉牛MSTN基因的遗传多态性及变异特征,采用PCR-SSCP方法检测了60头甘肃武威西门塔尔牛MSTN基因的多态性,对群体内各等位基因进行了克隆测序。结果显示：武威西门塔尔牛MSTN基因第1外显子存在A、B两个等位基因以及AA、AB两种基因型,基因型频率分别为0.9167、0.0833;第2外显子存在A、B两个等位基因以及AA、BB、AB三种基因型,基因型频率分别为0.5、0.25、0.25。序列分析表明,武威西门塔尔牛MSTN第1外显子在269 bp发生了A→G的突变,第2外显子在41 bp发生了C→T的突变,二者均属于同义突变。统计结果表明,武威西门塔尔牛MSTN基因第1外显子呈低度多态,第2外显子呈中度多态,外显子3没有发生突变。 相似文献
8.
[目的]为分析金昌地区西门塔尔牛 MSTN 基因三个外显子的遗传多态性及变异特征。[方法]采用 PCR-SSCP 方法检测了61头西门塔尔牛 MSTN 基因三个外显子的多态性。[结果]显示：金昌西门塔尔牛 MSTN 基因的第1外显子存在 A、B 两个等位基因以及AA、AB 两种基因型,其基因型频率分别为0.7705和0.2295;第2外显子存在 A、B 两个等位基因以及 AA、AB、BB 三种基因型,其基因型频率分别为0.0328、0.3443和0.6229;第3外显子只有 AA 一种基因型。序列分析表明,金昌地区的西门塔尔牛 MSTN 基因第1外显子在269bp 发生了 A→G 的突变和第2外显子在41bp 发生了 C→T 的突变,但都未导致氨基酸发生改变,属于同义突变;外显子3并未检测到突变。[结论]统计结果表明,金昌地区的西门塔尔牛 MSTN 基因第1外显子呈低度多态,第2外显子呈中度多态。 相似文献
9.
藏猪SLA-DRB基因第2外显子PCR-RFLP多态性分析 总被引：1，自引：0，他引：1
采用限制性内切酶RsaⅠ对藏猪SLA-DRB基因外显子2进行多态性分析。结果表明：藏猪SLA-DRB基因外显子2在RsaⅠ-RFLP位点上存在7种基因型，以杂合子BC型居多，基因型频率为0.4419，为优势基因型。χ2适合性检验结果表明，藏猪SLA-DRB基因外显子2 的RsaⅠ酶切位点的χ2值为10.14，高于显著水平（P<0.05），说明藏猪SLA-DRB基因在RsaⅠ酶切位点上没有达到Hardy-Weinberg平衡状态。 相似文献
10.
为了研究合作猪γ-IFN基因的多态性,确定其等位基因数、核苷酸多态位点,各等位基因是否符合Hardy-Weinberg平衡状态、多态信息含量、有效等位基因数和杂合度,采用PCR-SSCP方法检测了138头合作猪γ-IFN基因外显子4的等位基因,并克隆、测序群体内变异产生的各等位基因序列,就基因型和等位基因单倍型序列数据开展分析。序列比对分析发现5个核苷酸多态位点,其中5 284 bp(A→G)、5 341 bp(G→A)、5 371 bp(A→G)为新发现突变位点。试验群体发现4种基因型AA、AB、AC和AD,其中A等位基因频率为0.963 8,为优势等位基因。经χ2适合性检验,该群体处于Hardy-Weinberg平衡状态(P>0.05)。γ-IFN基因遗传多态指数中,杂合度(He)、多态信息含量(PIC)、有效等位基因数(Ne)均较低,表明合作猪γ-IFN基因比较保守。 相似文献
11.
采用PCR-RFLP技术研究了364头荷斯坦牛POU1 F1基因第二外显子的TaqⅠ酶切位点(e2-TaqⅠ)和第六外显子的HinfⅠ酶切位点(e6-HinfⅠ)多态性,并分析了两个位点多态性与泌乳量的关系。结果表明,实验群体中这两个多态位点均处于Hardy-Weinberg平衡状态,e2-TaqⅠ位点等位基因G/A的频率为0.2706/0.7294,e6-HinfⅠ位点等位基因G/A频率为0.3228/0.6772。性状关联结果表明,两个位点的基因型与第一、二胎泌乳量相关性不明显。 相似文献
12.
13.
D.C. Herrmann G. Wibbelt M. Götz F.J. Conraths G. Schares 《Veterinary parasitology》2013,191(1-2):108-111
Six free-ranging European beavers (Castor fiber) from Berlin greater metropolitan area and twelve European wildcats (Felis silvestris silvestris) originating from the German Federal State of Saxony-Anhalt were found dead and their carcasses were submitted for necropsy. Brain and lung samples were analysed for the presence of Toxoplasma gondii DNA. Histo-pathologic analysis of one beaver revealed several cyst-like protozoal structures in parts of the brain. Tissue DNA isolated from all animal samples was analysed by a specific T. gondii-PCR. Two beavers and four wildcats tested T. gondii-positive. DNA of the parasites was further analysed by PCR-RFLP typing using nine markers (nSAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico). Only T. gondii type II alleles were detected, except for the Apico locus, where type I alleles were observed in two isolates from beavers and in three from wild cats. The results of this study indicate that type II T. gondii (including type II variant strain) is the most common genotype infecting wildcats and beavers from Germany. 相似文献
14.
GUO Zhen-gang ZHANG Li-chun CAO Yang YU Yong-sheng LUO Xiao-tong JIN Hai-guo 《中国畜牧兽医》2013,40(10):184-188
In order to study the effect of myostatin (MSTN) gene on the meat performance of Local Yellow cattle in China,selecting 2 Local cattle breeds (Grassland Red bull,Yanbian cattle) as the research object,four binary hybrid cattle group (Germany Yellow cattle×Simmental,Limousin×Simmental,Charolais×Simmental and Red Angus×Simmental) as reference.PCR-RFLP and DNA sequencing methods were conducted to detect single nucleotide polymorphism of MSTN gene in two Chinese native cattle and four cross cattle,and analyzed the correlations between single nucleotide polymorphism of China Red Steppe MSTN gene and slaughter traits.The sequence analysis showed that there was no mutations in the MSTN gene exon 3,a C5358A mutation of 3'-UTR of MSTN gene was found,which made the AclⅠrestriction enzyme site disappeared,all cattle groups showed polymorphism which was formed two genes as A,C, and 3 genotypes as AA,CA and CC, respectively.The research results suggested that AA genotype in groups of four hybrid cattle had dominant advantage and the genotype frequency was apparently higher than two groups of Chinese Native cattle for the test of independence,allele A was expressed the preferential allelic,the frequencies of CA,CC and AA genotypes showed a little variation between two cross cattle and Native cattle,and the frequency of allele C was apparently higher than allele A,so allele C was designed the preferential allelic in the local population.While correlations between different genotypes of China Red Steppe and slaughter traits analysis showed that AA genotype individual net meat percentage and eye muscle area were significantly higher than CA and CC genotype individuals (P<0.05),but other slaughter straits had no significant differences in three genotypes(P>0.05).These results provided a useful reference for research of the meat performance of China Red Steppe in the molecular breeding field. 相似文献
15.
Allelic polymorphism in the ovine DQA1 gene 总被引：9，自引：0，他引：9
Variation in the ovine DQA1 gene was investigated by amplification of exon 2 using PCR, followed by single-strand conformational polymorphism (SSCP) analysis, cloning, and DNA sequencing. Fourteen novel SSCP patterns, representing 14 different sequences, were identified. Eight of these 14 sequences were identical to published DQA1 sequences from sheep, whereas the remaining six were novel but similar to the published DQA1 sequences from sheep and cattle. These six new sequences exhibited conserved region and variable region patterns similar to the published sheep DQA1 sequences, but were different than the published DQA2 sequences from sheep. All of these 14 putative sheep DQA1 sequences fulfilled the criteria used by the established bovine leukocyte antigens major histocompatibility complex nomenclature committee for assignment as new alleles. Comparison of the available DQA1 sequences from sheep and cattle revealed several clusters of ovine DQA1 sequences, and some sheep alleles were more similar to cattle alleles than other sheep alleles. The occurrence of trans-species polymorphism suggests the action of balancing selection at the DQA1 locus. Twenty-four percent of the nucleotide positions showed variation within exon 2, and this variation seems to have arisen largely by point mutation and gene conversion. The nonsynonymous and synonymous substitution rates were similar in both the putative antigen-binding site codons and the putative nonantigen-binding site codons. The extensive polymorphism reported in this article is consistent with polymorphism reported at the bovine DQA1 locus. 相似文献
16.
[目的]对武定鸡脂联素基因（NC_006096）第1外显子上的c.A99G突变位点进行多态性分析。[方法]翅静脉采集67只武定鸡全血,提取基因组DNA,采用PCR-RFLP法分析Hsp92Ⅱ限制性内切酶对PCR产物进行单酶切的结果。[结果]在c.A99G同义突变位点上存在AA、AB和BB 3种基因型,其中A基因频率为0.686 6,B基因频率为0.313 4;PIC=0.337 8,表明该位点为中度多态位点;χ2适合性检验表明,脂联素基因c.A99G位点处于平衡状态。[结论]采用PCR-RFLP法可有效检测武定鸡脂联素基因c.A99G突变,该位点有3种基因型,等位基因呈中度遗传多态水平且处于平衡状态。 相似文献
17.
马生长激素基因多态性与体尺指标之间的关联性分析 总被引：1，自引：0，他引：1
为研究广西德保矮马、百色马和新疆伊犁马的生长激素(GH)基因多态性与体尺指标之间的相关性,本实验利用PCR-SSCP技术分析3个品种277匹马GH基因5′侧翼区、第3外显子、第4外显子、第5外显子的遗传多态性。结果表明:只有GH基因第5外显子出现多态性,表现为AA、BB和AB 3种基因型,其他片段未发现多态性。群体遗传学分析表明,等位基因A和B的基因频率相等;Hardy-Weinberg平衡检验显示,3个品种马均处于非平衡状态;它们的多态信息含量均为中度多态。统计分析表明,GH基因第5外显子的基因型与3个品种马体尺指标之间无显著相关性。 相似文献
18.
本研究旨在利用高分辨率熔解曲线(high resolution melt,HRM)分型方法检测大白猪黑素皮质素受体(melanocortin-4 receptor,MC4R)和细胞分裂周期蛋白16(cell division cycle 16,CDC16)基因的多态性,并对其与大白猪的生长性状进行关联性分析。试验共采集246头大白猪的耳组织样提取基因组DNA,对小群体样本进行PCR扩增后,通过测序方法寻找两个基因在该群体中的多态性位点,针对已发现的多态性位点设计HRM专用引物,利用HRM方法对大群体进行多态性检测,将所得结果与大白猪生长性状进行关联性分析。结果表明,在MC4R基因序列中检测到1个突变位点2207A>G,2个等位基因:A和G,3个基因型:AA、AG和GG;在CDC16基因第18外显子上检测到了1个突变位点837T>C,2个等位基因:T和C,2种基因型:TT和CT。χ~2适合性检验结果显示,2个基因的基因型频率和等位基因频率在2个种群中分布差异不显著,符合Hardy-Weinberg平衡定律(P>0.05)。多态信息含量(PIC)分析显示,MC4R基因在2个群体中均处于中度多态(0.25相似文献
19.
采用PCR-RFLP方法对4个不同品种家兔MHC-DQA基因外显子2的遗传多态性进行检测,并进行聚类分析。结果表明,在家兔MHC-DQA基因外显子2的第103 bp处表现出多态性;χ2适合性检验结果表明,Mbo Ⅱ酶切位点在齐卡大型新西兰白兔和齐卡巨型白兔群体均没有达到Hardy-Weinberg平衡状态(P＜0.05),而在齐兴肉兔和加利福尼亚兔群体均达到Hardy-Weinberg平衡状态(P＞0.05);聚类分析结果表明,齐卡大型新西兰白兔与齐卡巨型白兔亲缘关系最近。试验结果为进一步利用MHC进行抗病育种提供了分子生物学基础。 相似文献
20.
Nicolaiewsky TB Richter MF Lunge VR Cunha CW Delagostin O Ikuta N Fonseca AS da Silva SS Ozaki LS 《Veterinary parasitology》2001,101(1):9-21
We describe a nested polymerase chain reaction (PCR) for the detection of Babesia equi in equine infected erythrocytes using oligonucleotides designed on the published sequence of a B. equi merozoite antigen gene (ema-1). A 102bp DNA fragment is specifically amplified from B. equi but not from Babesia caballi, Babesia bovis or Babesia bigemina DNA. In a mock infection we were able to detect down to six infected cells in 10(8) equine erythrocytes or to detect the parasite in blood with an equivalent parasitemia of 0.000006%. Furthermore, gene polymorphism was found by performing a PCR-RFLP (PCR combined with restriction fragment length polymorphism) on both the 102bp and the entire ema-1 gene DNA amplified from two B. equi isolates, Florida (USA) and Pelotas (Southern Brazil) isolates. The polymorphism was confirmed by sequencing the entire ema-1 gene from the B. equi isolate Pelotas. Our results demonstrate that the ema-1 based nested PCR is a valuable technique for routine detection of B. equi in chronically infected horses. It may be used for epidemiological and phylogenetic studies of the parasite as well as monitoring B. equi infected horses in chemotherapeutic trials. 相似文献