Xu A-Juan, Liu Xiao-Lin, Guo Jia-Zhong, Xia Zhi
Shaanxi Key Laboratory of Molecular Biology for Agriculture, Animal Science Technology College of Northwest A & F University, Yangling 712100, China.
Yi Chuan. 2010 Sep;32(9):929-34.
Four hundred and seven Chinese Holstein cows were chosen and divided into 2 groups (healthy group (317), sub-clinical and clinical group (100)) to investigate variation in bovine Tumor necrosis factor-alpha (TNF-a) gene and analyze their associations with mastitis. By PCR-RFLP and CRS-RFLP analyses, three polymorphic sites were detected, with one G→A mutation at 39 bp in exon2 and one C→T mutation at 293 bp in exon4, and C→G mutation in 5´UTR. The three polymorphic sites were able to be cut by DraⅠ, AfaⅠ, and DdeⅠ restriction enzymes respectively, and the DraⅠlocus was Create Restriction Enzyme Cutting Site. Chi-square analysis suggested that the three polymorphic sites did not reach Hardy-Weinberg equilibrium (P<0.05). Least square linear model (LSM) analysis indicated that the DraⅠ, AfaⅠ, and DdeⅠloci of TNF-a gene were associated with somatic cell score (SCS) (P<0.05). Genotype AA in DraⅠlocus and BB in AfaⅠlocus showed significantly higher SCS (P<0.05) and genotype AB in DdeⅠlocus showed significantly lower SCS than AA and BB genotypes in the population tested (P<0.05). This suggested that genotypes BB in DraⅠloci, AA in AfaⅠloci, and AB in DdeⅠloci can be used as candidate markers for mastitis resistance selection in dairy cattle.
选取407头中国荷斯坦奶牛,分为2组(健康组(317头)、亚临床和临床组(100头)),以研究牛肿瘤坏死因子-α(TNF-α)基因的变异情况,并分析其与乳腺炎的关联。通过PCR-RFLP和CRS-RFLP分析,检测到3个多态性位点,其中外显子2中39 bp处有一个G→A突变,外显子4中293 bp处有一个C→T突变,5´UTR中有一个C→G突变。这3个多态性位点分别可被DraⅠ、AfaⅠ和DdeⅠ限制性内切酶切割,DraⅠ位点为创建限制性内切酶切割位点。卡方分析表明,这3个多态性位点未达到Hardy-Weinberg平衡(P<0.05)。最小二乘线性模型(LSM)分析表明,TNF-α基因的DraⅠ、AfaⅠ和DdeⅠ位点与体细胞评分(SCS)相关(P<0.05)。DraⅠ位点的AA基因型和AfaⅠ位点的BB基因型显示出显著更高的SCS(P<0.05),DdeⅠ位点的AB基因型显示出比所测试群体中的AA和BB基因型显著更低的SCS(P<0.05)。这表明DraⅠ位点的BB基因型、AfaⅠ位点的AA基因型和DdeⅠ位点的AB基因型可作为奶牛抗乳腺炎选择的候选标记。