title: Novel SNPs in HSP70A1A gene and the association of polymorphisms with thermo tolerance traits and tissue specific expression in Chinese Holstein cattle.
abstract: Heat stress induces heat shock proteins (HSPs) expression and HSP70 family is one of them that have been reported to involve in cellular protection against heat stress. But whether there is any association of genetic variation in the HSP70A1A gene with thermo tolerance is unknown. PCR-SSCP and DNA sequencing were used to detect possible SNPs in HSP70A1A gene in 890 Chinese Holstein cattle. Three fragments were amplified and five novel mutations were found in HSP70A1A gene in Chinese Holstein cattle. G/A mutation was found at nucleotide 1524 in coding region that resulting two genotypes of AA and AB. T/C mutation was found at nucleotide 3494 in 3'-UTR resulting three genotypes of CC, CD and DD. The other three point mutations, G/C at nucleotide 6400, C/T at nucleotide 6600 and G/A at nucleotide 6601 were also found in 3'-UTR resulting six genotypes of EE, EF, FF, EG, FG and GG. Linkage disequilibrium (LD) analysis showed that the nucleotide 6400, 6600 and 6601 were in strong LD (D>0.75). Association analysis indicated that AB, DD and FF genotype were the thermo tolerance genotype. SBYE Green I was used to quantify HSP70A1A mRNA expression in different tissues through quantitative real-time PCR assay. The results of the real-time PCR showed that the expression of HSP70A1A mRNA in the heart was significantly higher than that in the other tissues (P<0.05). We presume that these mutations could be used in marker assisted selection for anti-heat stress cows in our breeding program.
authors.affiliation: Dairy Cattle Research Centre, Shandong Academy of Agricultural Science, Ji'nan, 250100, Shandong, People's Republic of China. liqiuling2000@yahoo.com.cn
authors.forename: Qiuling
authors.initials: Q
authors.lastname: Li
authors.forename: Jianbo
authors.initials: J
authors.lastname: Han
authors.forename: Fanglei
authors.initials: F
authors.lastname: Du
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Ju
authors.forename: Jinming
authors.initials: J
authors.lastname: Huang
authors.forename: Ji
authors.initials: J
authors.lastname: Wang
authors.forename: Rongling
authors.initials: R
authors.lastname: Li
authors.forename: Changfa
authors.initials: C
authors.lastname: Wang
authors.forename: Jifeng
authors.initials: J
authors.lastname: Zhong
mesh_terms: D000818:Animals; D001483:Base Sequence; D001947:Breeding / Q000379:methods*; D002417:Cattle / Q000235:genetics*; D002681:China; D015870:Gene Expression; D005819:Genetic Markers / Q000235:genetics; D055106:Genome-Wide Association Study; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D018869:Heat-Shock Response / Q000235:genetics*; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D018807:Polymorphism, Single-Stranded Conformational; D017422:Sequence Analysis, DNA
chemical_list: D005819:Genetic Markers; D018840:HSP70 Heat-Shock Proteins