Key Laboratory of Farm Animal Genetic Resources and Utilization of Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, People's Republic of China.
Mol Biol Rep. 2012 Mar;39(3):2909-14. doi: 10.1007/s11033-011-1052-3. Epub 2011 Jun 18.
In the present study, the exon 2 and 3' end sequence of intron 1 of BoLA-DRB3 gene of 211 Beijing Holstein cows was amplified and a uniform fragment of 284 bp was obtained. The genetic polymorphism was investigated by PCR-RFLP using two restriction endonucleases HaeIII and BstYI. Seven genotypes were detected by digesting the PCR products with HaeIII. The frequency of AA, AB, AC, AD, BB, BC and BF genotypes was 0.4638, 0.0193, 0.0193, 0.3478, 0.0290, 0.0386 and 0.0822, respectively. Three genotypes were found by digesting the PCR products with BstYI. The frequency of AA, AB and BB genotypes was 0.0569, 0.2844 and 0.6587, respectively. The relationship between the polymorphisms in exon 2 of BoLA-DRB3 gene and somatic cell score (SCS) in Beijing Holstein cows was analyzed by least squares linear model. No significant difference was detected among least squares means of SCS for seven HaeIII-RFLP genotypes (P > 0.05). As for BstYI-RFLP analysis, least squares mean of SCS for AA was significantly lower than that for AB (P < 0.05) or BB (P < 0.05), and no significant difference was detected between AB and BB genotypes (P > 0.05). BstYI AA was the most favorable genotype and BstYI BB was the most unfavorable genotype for mastitis resistance. The information found in the present study is very important for improving mastitis resistance in dairy cattle by marker assisted selection.
在本研究中，扩增了 211 头北京荷斯坦奶牛 BoLA-DRB3 基因第 2 外显子和第 1 内含子 3'端序列，获得了 284bp 的一致片段。采用 PCR-RFLP 法，用两种限制性内切酶 HaeIII 和 BstYI 研究了遗传多态性。通过 HaeIII 消化 PCR 产物检测到 7 种基因型。AA、AB、AC、AD、BB、BC 和 BF 基因型的频率分别为 0.4638、0.0193、0.0193、0.3478、0.0290、0.0386 和 0.0822。通过 BstYI 消化 PCR 产物发现 3 种基因型。AA、AB 和 BB 基因型的频率分别为 0.0569、0.2844 和 0.6587。采用最小二乘线性模型分析了 BoLA-DRB3 基因第 2 外显子多态性与北京荷斯坦奶牛体细胞评分（SCS）的关系。在 7 种 HaeIII-RFLP 基因型的 SCS 最小二乘均值之间未检测到显著差异（P > 0.05）。对于 BstYI-RFLP 分析，SCS 的最小二乘均值 AA 显著低于 AB（P < 0.05）或 BB（P < 0.05），AB 和 BB 基因型之间无显著差异（P > 0.05）。BstYI AA 是最有利的基因型，BstYI BB 是最不利于抗乳腺炎的基因型。本研究发现的信息对通过标记辅助选择提高奶牛乳腺炎抗性非常重要。
