摘要
以浙江省金华市伊康乳业奶牛场核心群357头中国荷斯坦牛为试验材料,旨在建立一种中国荷斯坦牛STAT4基因内含子20的单核苷酸多态性的简便检测方法。首先采用PCR-SSCP检测,结合DNA测序方法对STAT4基因内含子20的遗传多态性进行分析,并根据突变形式将PCR-SSCP检测方法转化为PCR-RFLP检测方法。结果发现STAT4基因内含子20内存在由g.95879 G>A和g.96013 G>C组成的完全连锁的单倍型模块。根据96013 G→C的突变可形成一个SduⅠ酶切多态位点的特性,建立了该单倍型的SduⅠ酶切检测体系。利用该体系检测到中国荷斯坦牛群体STAT4内含子20内存在GGGG,GAGC和AACC 3种不同的基因型,基因型频率分别为0.0784,0.4538和0.4678,而GG和AC基因频率分别为0.3053和0.6947。
This study was aimed to establish a simple detective method for polymorphisms of Chinese Holstein STAT4 gene intron 20. Three hundred and fifty-seven Chinese Holstein cows were used in this study. Firstly, the polymorphism of intron 20 of STAT4 gene was detected by PCR-SSCP method combined with DNA sequencing analysis. Then the detective method was transformed from PCR-SSCP to PCR-RFLP according to DNA saquencing results of the mutant SSCP patterns. The results showed that intron 20 of STAT4 gene had a complete linkage hap＇lotype block （g. 95879 G 〉 A and g. 96013 G 〉 C. A）. Sdu I enzyme detective system was established according to the characteristic that the 96013 G→C mutation may be formed as a restriction enzyme Sdu I polymorphism. By this detective system, three different genotypes, GGGG, GAGC and AACC were found, and the corresponding genotype frequencies were 0. 0784, 0. 4538 and 0. 4678. The frequencies of alleles GG and AC were 0. 3053 and 0. 6947, respectively.
出处
《浙江农业学报》
CSCD
北大核心
2012年第5期777-781,共5页
Acta Agriculturae Zhejiangensis
基金
浙江省重大科技专项(2008C02002-1
2010C12007)
国家科学技术部科技富民强县专项(国科发农[2010]349号)
