遗传 ›› 2010, Vol. 32 ›› Issue (9): 929-934.doi: 10.3724/SP.J.1005.2010.00929

• 研究报告 • 上一篇    下一篇

TNF-a基因多态性及其与奶牛乳房炎的相关性分析

徐阿娟, 刘小林, 郭家中, 夏志   

  1. 西北农林科技大学动物科技学院, 陕西省农业分子生物学重点实验室, 杨凌 712100
  • 收稿日期:2009-12-24 修回日期:2010-03-23 出版日期:2010-09-20 发布日期:2010-09-20
  • 通讯作者: 刘小林 E-mail:liuxiaolin@nwsuaf.edu.cn
  • 基金资助:

    国家十一五“863”项目(编号:2008AA10Z144), 陕西省“13115”科技创新项目(编号:2008ZDKG-11)和西安市科技计划项目 (编号:NC09049-2)资助

Polymorphism of bovine TNF-a gene and its association with mastitis in Chinese Holstein cows

XU A-Juan, LIU Xiao-Lin, GUO Jia-Zhong, XIA Zhi   

  1. Shaanxi Key Laboratory of Molecular Biology for Agriculture, Animal Science Technology College of Northwest A & F University, Yan-gling 712100, China
  • Received:2009-12-24 Revised:2010-03-23 Online:2010-09-20 Published:2010-09-20
  • Contact: LIU Xiao-Lin E-mail:liuxiaolin@nwsuaf.edu.cn

摘要: 以417头中国荷斯坦奶牛为研究对象, 根据体细胞评分(Somatic cell score, SCS)的大小将该奶牛群体划分为感染牛群(100头)和健康牛群(317头)。通过PCR-RFLP和CRS-RFLP方法检测了肿瘤坏死因子a(Tumor necrosis factor-alpha, TNF-a)基因在荷斯坦奶牛群体中的多态性, 并分析这些多态位点和奶牛乳房炎的相关性。研究发现了3个单核苷酸多态位点(Single-nucleotide polymorphism, SNP): 第2外显子39 bp处G→A的突变; 第4外显子293 bp处 C→T的突变; 5′侧翼区(5′-flanking region, 5′UTR)C→G的突变。这3个突变位点分别是DraⅠ、AfaⅠ和DdeⅠ限制性内切酶的酶切多态位点, 其中DraⅠ为创造酶切位点。经过基因型分析与χ2检验表明: 3个酶切多态位点在荷斯坦奶牛群中均未达到Hardy-Weinberg平衡状态。运用SPSS 13.0软件, 采用最小二乘拟合线性模型分析3个酶切多态位点与SCS的关系, 结果表明: AA基因型个体在DraⅠ酶切位点中的SCS显著大于BB及AB基因型个体(P<0.05), BB基因型表现出乳房炎抗性。AfaⅠ酶切位点中BB基因型个体的SCS显著大于AAAB基因型个体(P<0.05), AA基因型表现出乳房炎抗性。DdeⅠ酶切位点中, AB基因型个体的SCS显著低于AA基因型个体(P<0.05), AB基因型为优良基因型。因此BB、AA、AB基因型分别为DraⅠ、AfaⅠ、DdeⅠ酶切位点中的优良基因型, 可作为分子标记应用于奶牛乳房炎抗性筛选。

关键词: TNF-a基因, PCR-RFLP, 多态性, 乳房炎, 相关性

Abstract: 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 DdeⅠ loci of TNF-a gene were associated with somatic cell score (SCS) (P<0.05). Genotype AA in DraⅠlocus and BB in AfaⅠ locus 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.

Key words: TNF-a gene, PCR-RFLP, polymorphism, mastitis, relativity