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.
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
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.
[1] B1ockC J,Chase C,Hansen P J.Inheritance of resistance of bovinepreimplantation embryos toheat shock:Relative importance of thematernal versus paternal contribution[J].Mol.Repro,and Dev,2002,63:32-37.
[2] Pociot F,Ronningen K S,Nerup J.Polymorphic analysis of the human MHC-linked heat shock protein 70(HSP70-2)and HSP70-hom genesin insulin-dependent diabetes mellitus(IDDM)[J].Scand J Immunol993,38(5):491-495.
[3] Frederick M,Ausubel.Short Protocols in Molecular Biology.[M].New York:John Wiley and Sows,2001,36:3-27.
[4] 史彬林.红细胞钾作为乳牛耐热性选择指标的遗传可靠性研究[D].南京:南京农业大学991.
[5] Huang S Y,Chen M Y,Lin E C.Effects of single nucleotide polymorphisms in the 5'-flanking region of heat shock protein 70-2 gene on semen quality in boars[J].Anim Reprod Sci 2002,70:99-109.
[6] Huang S Y.Lee W C.Chen M Y.Genotypes of 5'-flanking region in porcine heat-shock protein 70.2 gene affect backfat thickness and growth performance in Duroc boars[J] Livest Prod Sci,2004 85:181-187.
[7] Elvinger F,Natzke R P,Hansen P J.Interactions of heat stress and bovine somasotropin affecting physiology and immunology of lactating cows[J].Dairy Science992,75:499-463.
[8] 穆玉云,李如治,黄昌澍.乳牛耐热性指标的检测[J].安徽农业大学学报993(1):66-71.
[9] Evans J V.Electrolyte concentrations in red blood cells of british breeds of sheep[J].Nature95474,931-932.
[10] Evans J V,King J W.Genetic control of sodium and potassium concentrations in the red blood cells of sheep[J].Nature95576(4473):171.
[11] Evans J V.Water metabolism in the sheep[J].Nature95780(4589):756.
[12] 史彬林,闫素梅.奶牛耐热性评定指标的研究[J].中国奶牛996(2):20-22.
