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1.
旨在运用多座位外显方差分析法(MPVA)分析中国荷斯坦牛CXCR1和IL8两个基因多个突变位点多态性与乳房炎易感性的交互作用.本研究以南方某大型奶牛场634头(其中102头患乳房炎)中国荷斯坦牛为试验材料,采用PCR-SSCP和直接测序法分析CXCR1-5′端和编码区及IL8 2789/2862内含子3、外显子4和5,2个基因共7个SNPs位点的遗传多态性,并用MPVA法分析CXCR1和IL8基因各位点突变及其基因型组合对奶牛乳房炎易感性的影响.结果,MPVA分析发现CXCR1-1830、CXCR1-1768和IL8 2789/2862三位点交互作用对奶牛乳房炎易感性的影响达到显著水平.CXCR1-1830、CXCR1-1768和IL8 2789/2862三位点交互模型作为奶牛乳房炎易感性相关的基因联合选择模型最佳,MPVA可用于奶牛乳房炎易感性多基因分析模型.  相似文献   

2.
牛整合素β-2(ITGB2)基因D128G突变能引起牛白细胞粘附综合征(BLAD),是牛乳房炎抗性的研究热点之一,但ITGB2基因其他突变在中国荷斯坦牛中的分布及其与乳房炎抗性和产奶性状是否有关尚不清楚。本研究选取ITGB2基因389CT(rs108953545)和774CT(rs209411752)2个SNP位点,利用SNaPshot技术检测其在1 212头荷斯坦母牛的基因型,并与体细胞评分(SCS)和5个产奶性状(305 d产奶量、乳脂和乳脂率、乳蛋白和乳蛋白率)进行相关性分析。结果表明:389位点CC、CT、TT基因型的频率分别是0.29、0.49和0.22,774位点相应的基因型频率为0.69、0.27、0.04;2个位点与SCS和5个产奶性状相关性分析显示,仅774位点CT和TT型个体乳蛋白率显著低于CC基因型(P0.05);2个位点6种复合基因型相关分析发现,CTCC和TTCC基因型具有高的乳脂率和乳蛋白率(P0.01),而TTCC基因型个体的SCS更低(P0.05)。总体而言,TTCC基因型是中国荷斯坦奶牛乳房炎抗性和乳品质潜在的分子标记。  相似文献   

3.
[目的]本研究旨在研究中国荷斯坦牛IL8基因-233(G>A)位点不同基因的表达差异水平,为探讨该位点在抗奶牛乳房炎作用提供理论依据.[方法]本试验运用SYBRGreen实时荧光定量PCR技术进行定量测定.[结果]GG基因型个体mRNA的表达量显示高于GA和AA基因型个体的表达量.[结论]IL8基因-233(G>A)位点对中国荷斯坦牛乳房炎抗性有一定影响,可用于中国荷斯坦牛的抗乳房炎的分子标记辅助选择.  相似文献   

4.
利用PCR-RFLP方法检测TLR2基因在297头中国荷斯坦牛中的多态性,并分析其多态性与体细胞评分(SCS)的相关性.结果显示,TLR2基因exon2扩增片断的109 bp处产生C→A的突变,并导致天冬氨酸改变为谷氨酸.TLR2基因exon2被EcoRV消化后表现多态性,表现AA、AB、BB 3种基因型,其中B等位基因为优势等住基因,等位基因频率为0.784 5,而A等位基因频率则为0.215 5.经X2适合性检验,中国荷斯坦牛在该位点达到Hardy-Weinberg平衡状态.同时,群体EcoRV基因座不同基因型与SCS相关分析的结果表明,BB、AB型个体的SCS指标显著高于AA型(P<0.05),AA基因型可作为乳腺炎抗性的有利基因型,可将TLR2作为奶牛乳腺炎候选基因,应用于乳腺炎抗性分子标记辅助选择育种.  相似文献   

5.
试验旨在研究中国荷斯坦牛Toll样受体4(toll-like receptor 4,TLR4)基因的遗传多态性及其与体细胞评分(somatic cell score,SCS)的关联性,寻找与乳房炎相关的分子标记,加快中国荷斯坦牛的抗病育种。利用PCR-RFLP和PCR-SSCP技术对30个公牛家系的610头中国荷斯坦牛TLR4基因进行多态性检测,用最小二乘均数法对TLR4基因的多态位点与SCS进行相关分析。结果发现,试验共检测到2个单核苷酸多态(single nucleotide polymorphisms,SNPs),TLR4基因5'侧翼区存在SNP -226 G>C突变,经PCR-RFLP检测发现3种基因型:GG、GC和CC,基因型频率分别为0.208、0.482和0.310;外显子3存在SNP 1760 C>T突变,经PCR-SSCP检测发现3种基因型:CC、TC和TT,基因型频率分别为0.738、0.225和0.007,以上2位点均偏离Hardy-Weinberg 平衡。对于SNP -226 G>C位点,基因型个体的SCS差异不显著;对于SNP 1760 C>T位点,CC基因型个体的SCS最小二乘均值极显著低于TT和TC基因型个体(P<0.01)。SNP 1760 C>T的CC基因型对于中国荷斯坦牛的SCS有较大的遗传效应,可作为分子标记应用于奶牛乳房炎抗性筛选。  相似文献   

6.
牛TLR4基因的遗传多态性与乳房炎的关联分析   总被引:10,自引:2,他引:10  
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共397头为研究对象,利用创造酶切位点PCR法扩增243bp的目的片段,通过限制性内切酶HinfⅠ酶切来检测TLR4第3外显子的多态性,结果发现扩增产物的27bp处C到T的突变使得多态位点产生,编码的氨基酸由苏氨酸变为异亮氨酸。A、B2个等位基因在3个群体中均有分布,A等位基因占优势(大于78%),经χ^2适合性检验,三河牛在该位点未达到Hardy-Weinberg平衡状态(P〈0.05)。运用SAS8.2软件采用最小二乘法拟合线性模型,将该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: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。  相似文献   

7.
以北京地区11个牧场的494头中国荷斯坦牛为试验材料,采用病例-对照(case-control)设计,研究了motilin(MLN)基因第一外显子内转录因子结合位点g.90TC多态性与奶牛真胃变位易感性的关联。结果共检测到TT、TC、CC三种基因型,基因型频率分布符合Hardy-Weinberg平衡(P0.05)。将该位点3种基因型与真胃变位易感性进行关联分析,结果发现在一胎牛中关联显著(Cochran-Armitage趋势检验P0.05;卡方检验P=0.09);与基因型TT相比,基因型TC和CC的比值比(OR值)分别为1.52和2.06,即易感性CCTCTT,显示C等位基因更易导致真胃变位。这些结果支持了MLN基因与荷斯坦牛真胃变位可能存在密切的关系。对g.90TC位点T等位基因的选择,将有利于提高群体对真胃变位的遗传抗性。  相似文献   

8.
通过研究奶牛溶菌酶基因(Bos taurus 1ysozyme,LYZ)的多态性与乳房炎的相关性,为奶牛的抗病育种提供新的分子遗传标记。利用PCR-SSCP技术对420头中国荷斯坦牛LYZ基因进行多态性检测,并对LYZ基因的多态位点与SCS(somatic cell score)和泌乳性状进行相关分析。结果表明:LYZ基因的外显子1上存在115(TG)突变位点,使精氨酸变为亮氨酸;发现了3种基因型GG,TG和TT,频率分别为0.20、0.48和0.32;GG基因型个体的SCS最小二乘均值极显著低于TT基因型个体(P0.01),显著低于TG基因型个体(P0.05);GG基因型个体的305d产奶量最小二乘均值极显著高于TT基因型个体(P0.01),显著高于TG基因型个体(P0.05)。LYZ基因该位点的突变对奶牛的SCS和305d产奶量有较大的遗传效应,可作为分子标记应用于奶牛乳房炎抗性和产奶量的筛选。  相似文献   

9.
牛BoLA-DRB3基因的多态性与乳房炎相关性初探   总被引:9,自引:0,他引:9  
利用PCR—RFLP技术对易感乳房炎的奶牛血样24份,不易感染乳房炎的2个黄牛品种南阳牛、秦川牛血样66份共计90个个体进行牛BoL A—DRB3基因第2外显子多态性分析。结果表明:扩增大小为252bp的片段经限制性内切酶BstUI酶切后表现多态,且奶牛AA型个体显著高于黄牛品种,提示BoLA等位基因A作为母牛乳房炎易感性的候选标记具有潜在的有效性。  相似文献   

10.
旨在分析奶牛金黄色葡萄球菌(金葡菌)抗性候选基因TRAPPC9的遗传效应。本研究利用中国北方地区6个牛场517头具有完整DHI、年龄及胎次记录的中国荷斯坦牛的生产数据,使用Multiplex SnaPshot微测序技术分析TRAPPC9基因4个SNPs的基因型,利用特异性PCR技术检测牛只感染金葡菌情况,同时测定血清中TNF-α、IFN-γ、IL-17、NF-κB等细胞因子的含量。在此基础上,利用GLM模型最小二乘法分析TRAPPC9基因4个SNPs对奶牛金葡菌乳房炎抗性的遗传效应。结果表明:IL-17、TNF-α和IFN-γ可作为有效的乳房炎参考指标。对于金葡菌阴性牛,TRAPPC9基因SNP4(C2477531T)对SCS有极显著影响(P0.01);对于金葡菌阳性牛,该SNP对IL-17有极显著效应(P0.01)。TRAPPC9基因可作为中国荷斯坦牛金葡菌乳房炎抗性分子标记之一。  相似文献   

11.
Holstein Cows ( n  = 702) from 26 dairy herds in the Tama area of Tokyo, Japan were examined for polymorphisms of the BoLA-DRB3 allele using a PCR-RFLP method. Twenty alleles were observed and allelic frequencies ranged from < 1% to 20.3%. Nine alleles ( DRB3.2 * 24, * 16, * 8, * 23, * 22, * 3, * 11 , * 10 and * 7 in order) constituted 90.0% of all alleles. Somatic cell counts (SCC) were used to classify healthy (group 1), mastitis (group 2) and suspected (group 3) cows. Frequencies of DRB3.2 * 11 and DRB3.2 * 23 were slightly higher in group 1 than in group 2, whereas, frequencies of DRB3.2 * 8 and DRB3.2 * 16 were slightly higher in group 2 than in group 1. However, none of the differences in frequencies between the two groups were statistically significant. For combinations of alleles, frequencies of DRB3.2 * 8/ * 23 ( P  < 0.1) and DRB3.2 * 16/ * 24 ( P  < 0.05) were significantly higher in group 2 than in group 1, and their odds ratios were 2.1, 2.5, respectively. However, there were no significant differences between genotypes in their effects for SCC. On the other hand, frequency of DRB3.2 * 23/ * 23 including combinations of DRB3.2 * 23 with minor alleles was significantly higher in group 1 than in group 2 ( P  < 0.01), and the odds ratio was 0.3. Therefore, it was considered that mastitis resistance or susceptibility of cows may vary with the combination of BoLA-DRB3 alleles.  相似文献   

12.
Bovine leukocyte antigen DRB3 alleles from Holstein x Zebu crossbred dairy cows (n = 409) were analyzed using the PCR-RFLP technique. Exon II of DRB3 was amplified using locus-specific primers (HLO30/HLO32), followed by digestion with 3 restriction enzymes (RsaI, BstyI, and HaeIII). Forty alleles were found with frequency ranging from 0.005 to 0.139. The most frequently detected alleles of Holstein x Zebu were DRB3*16, *51, *23, *11, *8, and *1, accounting for 61.12% of the alleles in the population. Detection of candidate alleles for clinical mastitis occurrence was performed by logistic regression. It was found that percentage of Holstein fraction in crossbred cows had a nonsignificant effect (P > 0.05). However, parity had a significant effect on mastitis occurrence. In addition, DRB3*1 and *52 were the most associated with the occurrence of clinical mastitis, whereas *15, *51, and *22 were associated with resistance in crossbred populations. This is the first report of association of DRB3*15 and *51 with mastitis resistance. The association was validated by examining the candidate alleles in another commercial population. Highly susceptible (n = 43) and resistant (n = 42) groups of Holstein x Zebu cows were investigated. The result confirmed that DRB3*1 and *52 could be considered as susceptibility alleles, whereas *15, *51, and *22 could be considered as resistant alleles in Holstein x Zebu raised under tropical conditions. In addition, allele effects on 305-d milk production were estimated by BLUP. It was shown that most alleles associated with high clinical mastitis occurrence were related to increased milk yield. This study revealed that allele DRB3*10 had the greatest effect on increasing milk yield with moderate resistance to clinical mastitis, which could be used as a potential marker for selection in dairy genetic evaluation.  相似文献   

13.
The association of the polymorphism of bovine leukocyte antigen ( BoLA-DRB3 ) genes identified by the polymerase chain reaction sequence-based typing (PCR-SBT) method with resistance and susceptibility to mastitis caused by pathogenic bacteria was investigated. Blood samples for DNA extraction were collected from 194 Holstein cows (41 healthy cows and 153 mastitis cows including 24 mixed-infection cows infected with 2 or 3 species of pathogens) from 5 districts of Chiba prefecture, Japan. Sixteen BoLA-DRB3 alleles were detected. The 4 main alleles of DRB3*0101 , *1501 , *1201 , and *1101 constituted 56.8% of the total number of alleles detected. Mastitis cows were divided into 2 groups: group 1 with single-infection cows and group 2 with all mastitis cows including 24 mixed-infection cows. The differences in the frequencies of BoLA-DRB3 alleles and the number of cows homozygous or heterozygous for each BoLA-DRB3 allele between healthy cows and the 2 groups of mastitis cows were evaluated. Furthermore, similar comparisons were performed between healthy cows and the 2 groups of mastitis cows for each mastitis pathogen. It was considered that the 4 alleles, namely, DRB3*0101 , *1501 , *1201 , and *1101 had specific resistance and susceptibility to 4 different mastitis pathogens. Thus, DRB3*0101 might be associated with susceptibility to coagulase-negative Staphylococci and Escherichia coli , and DRB3*1501 might be associated with susceptibility to Escherichia coli . However, DRB3*1101 might be associated with resistance to Streptococci and coagulase-negative Staphylococci , and DRB3*1201 , with resistance to Streptococci , Escherichia coli , and Staphylococcus aureus .  相似文献   

14.
The major histocompatibility complex (MHC) is a gene complex closely linked to the vertebrate immune system due to its importance in antigen recognition and immune response to pathogens. To improve our understanding of the MHC and disease resistance in dairy cattle, we gathered 5119 test day records of somatic cell count (SCC) and performance traits of 262 Holstein dairy cows to determine whether the DRB region of the MHC contains alleles that are associated with elevated SCC, milk yield, protein and fat percent of milk. To this purpose, genotyping of animals for DRB3 gene was investigated by polymerase chain reaction‐based restriction fragment length polymorphism (PCR‐RFLP) assay. A two‐step PCR was carried out so as to amplify a 284 base‐pair fragment of exon 2 of the target gene. Second PCR products were treated with three restriction endonuclease enzymes RsaI, BstYI and HaeIII. Twenty‐eight BoLA‐DRB3 alleles were identified including one novel allele (*40). The results in general are in good accordance with allele frequencies of Holstein cattle populations reported by previous studies. Analyses of associations were modeled based on repeated measurement anova and generalized logistic linear methods for production traits and SCC data, respectively. The results of this study showed a significant relationship between the elevated SCC reflecting an increased probability of occurrence to subclinical mastitis and DRB3.2 allele *8 (p < 0.03). The results also revealed significant positive relationships of alleles*22 (p < 0.01) and allele*11 (p < 0.05) with milk fat percent as well as of alleles*24 (p < 0.03) and *22 (p < 0.05) with protein percent. The present study failed to find any association between milk yield and tested alleles. Because of the lack of consistency among results of similar studies, we suggest further investigations to determine the precise nature of these associations with the high polymorphic bovine MHC region to be performed based on haplotypes.  相似文献   

15.
【目的】 探究磷脂酰肌醇-4,5-二磷酸3-激酶催化亚单位β(PIK3CB)基因多态性及其与中国荷斯坦牛繁殖和产奶性状的关系。【方法】 通过混池测序对中国荷斯坦牛PIK3CB基因进行单核苷酸多态性(SNP)位点筛选,采用竞争性等位基因特异性PCR (KASP)技术在1 160头健康泌乳中国荷斯坦牛中进行SNP分型并进行群体遗传学分析,采用线性模型进行SNP与11个繁殖和产奶性状基于单位点和单倍型组合的关联分析。【结果】 在PIK3CB基因中共检测到了17个SNPs,筛选出7个SNPs用于后续分析。关联分析发现,7个SNPs与多个目标性状存在显著或极显著的关联(P<0.05;P<0.01);位于外显子区域的g.130433743 A>G位点AA基因型个体和位于可变剪接区域的g.130448069 G>A位点GG基因型个体,其经产牛首末次配种间隔、产奶量、乳蛋白量和乳脂量最低,体细胞评分最高,上述基因型个体具有较短的首末次配种间隔,而产奶性能相对较差;g.130387717 G>A位点AA基因型个体,其初配日龄和青年牛首末次配种间隔最低,产奶量、乳蛋白量和乳脂量最高,该基因型个体的繁殖和产奶性能均较好,上述3个SNPs位点可作为中国荷斯坦牛繁殖和产奶性状的候选位点重点关注。单倍型分析发现,PIK3CB基因的g.130387717 G>A、g.130430832 A>-、g.130433743 A>G、g.130433982 C>T、g.130446073 C>T和g.130448069 G>A 6个SNPs紧密连锁形成一个单倍型块,且与多个目标性状存在显著或极显著关联(P<0.05;P<0.01),其中H2H3和H2H4单倍型组合个体的繁殖和产奶性能较好,为优势单倍型组合。【结论】 中国荷斯坦牛PIK3CB基因存在丰富的遗传变异,其多态性与繁殖和产奶性状存在关联,g.130433743 A>G、g.130448069 G>A和g.130387717 G>A位点可作为潜在分子标记,为中国荷斯坦牛的平衡育种提供理论依据。  相似文献   

16.
通过研究中国荷斯坦牛趋化因子受体1(Chemokine(C-X-C motif)receptor1,CXCR1)基因多态性与乳腺炎间的相关性,找到对乳腺炎性状有显著影响的基因型和单倍型组合,为从遗传角度上解决奶牛乳腺炎问题提供参考。采用巢氏PCR、DNA测序和CRS-PCR-RFLP方法,对中国荷斯坦牛的CXCR1外显子2进行遗传多态性研究,分别利用SHEsis软件和PHASE软件进行配对连锁不平衡分析和单倍型分析。结果表明,找到了4个新SNPs,分别为291(C/T)、333(C/T)、337(A/G)和365(C/T)。各SNPs与中国荷斯坦牛体细胞评分(SCS)和产奶量的关联分析表明:337(A/G)和365(C/T)位点的等位基因G、C均为低SCS、高产奶量的优秀等位基因;另外,H3 H7(CCCTGGCC)单倍型杂合个体为低SCS、高产奶量的优秀单倍型组合。CXCR1基因的单倍型H3 H7(CCCTGGCC)在SCS和产奶量方面是优良的单倍型,可作为选择抗乳腺炎的分子标记。  相似文献   

17.
In order to study mutations of bovine breast cancer 1 (BRCA1) gene and its association with somatic cell count and somatic cell score in Holstein dairy cows in Beijing.SNPs of BRCA1 gene exon 13 and 14 were obtained by cloning,sequence alignment and searching for known mutations.In our research,25025 T>A of exon 9 and 46126 G>T of exon 13 in BRCA1 were detected by SNaPshot in Holstein dairy cows raised on the outskirts of Beijing city,as well as the association between SNPs and somatic cell count (SCC) or somatic cell score (SCS) were also analysed.The results showed that both 25025 T>A and 46126 G>T were detected in Holstein dairy cows.TT genotype at locus 25025 T>A and GT genotype at locus 46126 G>T were dominant genotypes,respectively.At locus 25025 T>A,least squares mean of SCC and SCS in individuals with AA genotype was significantly lower than that with TT and TA genotypes (P<0.05;P<0.01).Least squares mean of SCC in individuals with TT genotype was significantly lower than that with GG and GT genotypes (P<0.05),while SCS of different genotypes showed no difference at locus 46126 G>T (P>0.05).These results preliminarily indicated that the 25025 T>A and 46126 G>T mutations of BRCA1 gene could be used in molecular marker-assisted selection programs of mastitis resistance in Holstein dairy cows.  相似文献   

18.
Genotyping of bovine leucocyte antigen DRB3.2 (BoLA-DRB3.2) in a total of 523 Norwegian Red (NR) cows from two groups selected for high protein yield and low clinical mastitis, respectively, identified 27 previously reported BoLA-DRB3.2 alleles across the groups. Significant differences in BoLA-DRB3.2 allele frequencies were found between the selection groups. Alleles *13, *18, *22 and *27 had a significantly higher frequency in cows selected for low clinical mastitis, while alleles *3, *9, *11 and *26 had a higher frequency in cows selected for high protein yield. Associations between BoLA-DRB3.2 alleles and clinical mastitis were analysed based on mastitis data from 741,072 first-lactation NR cows, of which 452 were genotyped. Alleles *22 and *26 were found to be associated with increased clinical mastitis, while alleles *7, *11, *18 and *24 had a favourable effect on mastitis resistance. Contradictory results from different studies investigating associations between BoLA-DRB3.2 alleles and mastitis indicate that future studies should focus on associations of mastitis with BoLA haplotypes rather than with single BoLA genes.  相似文献   

19.
为了研究BRCA1基因突变与荷斯坦奶牛体细胞数和体细胞评分的关系,试验通过对BRCA1基因外显子13、14进行克隆、序列比对和挖掘已有突变的方法确定该基因的多态位点,采用SNaPshot技术检测了BRCA1基因25025 T>A和46126 G>T突变位点在北京郊区荷斯坦奶牛群体中的分布,并对突变位点与体细胞数和体细胞评分进行了关联分析。结果表明,荷斯坦奶牛BRCA1基因2个位点均检测到3种基因型,其中25025 bp位点TT基因型为优势基因型,46126 bp位点GT基因型为优势基因型。25025 bp位点AA基因型个体体细胞数(P<0.05)和体细胞评分(P<0.01)都显著低于TT和TA基因型;46126 bp位点TT基因型个体体细胞数显著低于GG和GT基因型个体(P<0.05),但3种基因型个体体细胞评分无显著差异(P>0.05)。本研究结果初步表明,BRCA1基因25025和46126 bp位点可作为中国荷斯坦牛乳房炎抗性的标记辅助选择。  相似文献   

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