摘要： 研究中国荷斯坦牛HSP70-1基因与奶牛耐热性之间的关系.根据NCBI登陆号为AY149618(19 26 bp)的HSP70-1基因序列设计9对引物分段扩增该基因全长,并用PCR-SSCP、PCR-RFLP和CRS-RFLP相结合的方法寻找中国荷斯坦牛HSP70-1基因编码区的多态位点,分析山东省3个现代化牛场产奶牛127头,并以直肠温度和红细胞钾离子浓度为荷斯坦牛的耐热指标,用SAS 软件最小二乘分析一般线性模型分析试验牛HSP70-1基因型与耐热指标的相关性.发现1 623 bp处发生G→A→C的复等位突变位点,2 409 bp处发生G→A的突变;χ2检验结果表明在试验牛群中两位点均未达到Hardy-Weinberg平衡;2409位点基因型与两个耐热指标相关性不显著(P＞0.0)623位点基因型与热应激时期中国荷斯坦牛的直肠温度相关性显著(P＜0.0),GA基因型个体显著低于CC和GG基因型个体,GA基因型为耐热基因型.HSP70-1基因可用作中国荷斯坦牛选育耐热牛群的分子遗传标记.
关键词:
热休克蛋白70,
限制性片段长度多态性,
单链构象多态性,
耐热性,
直肠温度
Abstract: Nine pairs of primer were designed according to the HSP7021 gene sequence of NCBI with AY149618 (19 526 bp) accession number. In combination of the PCR2SSCP,PCR2RFLP and CRS2RFLP method,the author tries to find out the polymorphic loci in gene coding region of Chinese Holstein Cattle HSP7021. It is found out that there was G2 A2C multiple allele in mutation site of 1 623 bp position and there was mutation of G2A in 2 409 bp position. The author also analyzed 127 milking cows from three modern cattle farms in Shandong Province. Theχ2 test resulted manifests that the two sites fails to reach the Hardy2Weinberg balance. The rectal temperature and the potassium concentration of ery2 throcyte were the heat resistance indexes of Holstein Cattle. The correlation between the genotype and the heat resistance index of HSP7021 experimental cattle was analyzed by least square analysis of SAS software and general linear model analysis. The result showed that the correlation between the genotype in 2 409 position and the two indexes was not obvi2 ous. The correlation between genotype in 1 623 position and the rectal temperature of the Chinese Holstein Cattle HSP702 1 in heat stress period was obvious ( P < 0. 05). The genotype birds of GA were obviously lower than that of CC and GG. GA genotype belongs to the heat resistance genotype. It reveals that HSP7021 gene could be used as the molecular genetic marker to breeding the heat assistance cattle for Chinese holstein cattle.
Key words:
Chinese holstein cattle HSP70,
Restriction fragment length polymorphism,
Single2strand conformation polymorphism,
Thermotolerance,
Rectal temperature
中图分类号:
程维杰, 李秋玲, 孙延鸣, 王洪梅, 王长法, 李建斌, 仲跻峰. 中国荷斯坦牛HSP70-1基因遗传多态性与其耐热性的关系分析[J]. 华北农学报, 2009, 24(4): 41-45. doi: 10.7668/hbnxb.2009.04.009.
CHENG Wei-jie, LI Qiu-ling, SUN Yan-ming, WANG Hong-mei, WANG Chang-fa, LI Jian-bin, ZHONG Ji-feng. The Relation Analysis between the Genetic Polymorphism and the Heat Resistance of Chinese Holstein Cattle HSP70-1[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2009, 24(4): 41-45. doi: 10.7668/hbnxb.2009.04.009.