摘要： 用PCR-SSCP和测序技术研究了541头中国荷斯坦奶牛热休克蛋白70(HSP70)基因的3’-非翻译区的多态性,并研究了多态性对奶牛产奶性状和耐热性状的影响。设计引物扩增片段大小549 bp,通过PCR-SSCP技术发现参与试验的奶牛出现了3个复等位基因:A、B和C,6种基因型:AA、AB、AC、BB、BC和CC基因型。测序发现复等位基因是由3个新的突变引起的,第6 400、6 600和6 601位碱基分别发生了G/C、C/T和G/A突变。χ2适合性检验表明,该群体在这三个位点的突变没有达到Hardy-Weinberg平衡状态(P<0.01)。中国荷斯坦牛BB基因型个体红细胞钾含量和产奶量下降率显著低于其他基因型个体(P<0.05),表明BB型的中国荷斯坦牛耐热性能优于其他类型。BB基因型个体305 d产奶量高于其他5种基因型的个体,进一步说明BB基因型可能在选育高产抗热应激奶牛中得到应用。
关键词:
荷斯坦奶牛,
HSP70基因,
多态性,
热应激,
PCR-SSCP
Abstract: Heat shock protein 70( HSP70) gene was regarded as a candidate gene for thermo resistance of cows.In this study,a 549 bp fragment of HSP70 gene of Chinese Holstein was amplified by polymerase chain reaction( PCR), the polymorphisms of three loci of HSP70 gene were detected by PCR-SSCP technique.By sequencing, three point mutations, G/C at nucleotide 6 400,C/T at nucleotide 6 600 and G/A at nucleotide 6 601,were identified in HSP70 gene sequence.χ2 test indicated that three polymorphisms in the population unfitted with Hardy-Weinberg equilibrium( P <0.01).Meanwhile, the relationship between milk performance and heat tolerance was analyzed.The results showed that in the three loci, the different genotypes of HSP70 gene had no significant influence on milk protein percent and fat percent,but had significant influence on 305-day milk yield.Individuals with genotype BB showed higher 305-day milk yield than those with genotypes AA and CC( P <0.05).And the cows with genotype BB showed lower potassium content in erythrocytes ( PCE) and decrease rate of milk yield than those with genotypes AA and CC.It indicated that BB genotype was the advantageous genotype.These mutations could be used in MAS for anti-heat stress cows in our breeding program．
Key words:
Holstein cows,
HSP70 gene,
Polymorphism,
Heat stress,
PCR-SSCP
中图分类号:
杜方磊, 王洪梅, 黄金明, 李建斌, 仲跻峰, 张廷荣, 刘剑飞, 李秋玲, 王长法. 中国荷斯坦牛HSP70基因3＇-非翻译区多态性与耐热性的关联分析[J]. 华北农学报, 2010, 25(3): 14-18. doi: 10.7668/hbnxb.2010.03.004.
DU Fang-lei,, WANG Hong-mei, HUANG Jin-ming, LI Jian-bin, ZHONG Ji-feng, ZHANG Ting-rong, LIU Jian-fei, LI Qiu-ling, WANG Chang-fa. Polymorphism at 3'UTR of the Heat Shock Protein 70 Gene and Its Relationship with Thermal Tolerance in Chinese Holstein[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2010, 25(3): 14-18. doi: 10.7668/hbnxb.2010.03.004.