Polymorphism of the Promoter Region of Hsp70 Gene and Its Relationship with the Expression of HSP70mRNA, HSF1mRNA, Bcl-2mrna and Bax-AMrna in Lymphocytes in Peripheral Blood of Heat Shocked Dairy Cows원문보기
The blood samples were collected from dairy cows at the same milking stage. The single-strand conformation polymorphism (PCR-SSCP) method was used to analyze for polymorphism at the 5'flanking region of the hsp70 gene. The mRNA expression levels of HSP70, HSF1, Bcl-2 and Bax-$\alpha$ at d...
The blood samples were collected from dairy cows at the same milking stage. The single-strand conformation polymorphism (PCR-SSCP) method was used to analyze for polymorphism at the 5'flanking region of the hsp70 gene. The mRNA expression levels of HSP70, HSF1, Bcl-2 and Bax-$\alpha$ at different daily-mean-temperature were analyzed by relative quantitative RTPCR. The DNA content, cell phase and the ratio of apoptosis of lymphocytes in peripheral blood of dairy cattle at different daily-meantemperature were determined by FCM. The PCR-SSCP products of primer pair 1 showed polymorphisms and could be divided into four genotypes: aa, ab, ac, cc, with the cis-acting element (CCAAT box) included. Mutations in the hsp70 5'flanking region (468-752 bp) had different effects on mRNA expression of HSP70, HSF1, Bcl-2 and Bax-$\alpha$. The ac genotypic cows showed higher expressions of HSP70mRNA, HSF1mRNA and Bcl-2mRNA/Bax-$\alpha$mRNA and lower ratio of apoptosis. These mutation sites can be used as molecular genetic markers to assist selection for anti-heat stress cows.
The blood samples were collected from dairy cows at the same milking stage. The single-strand conformation polymorphism (PCR-SSCP) method was used to analyze for polymorphism at the 5'flanking region of the hsp70 gene. The mRNA expression levels of HSP70, HSF1, Bcl-2 and Bax-$\alpha$ at different daily-mean-temperature were analyzed by relative quantitative RTPCR. The DNA content, cell phase and the ratio of apoptosis of lymphocytes in peripheral blood of dairy cattle at different daily-meantemperature were determined by FCM. The PCR-SSCP products of primer pair 1 showed polymorphisms and could be divided into four genotypes: aa, ab, ac, cc, with the cis-acting element (CCAAT box) included. Mutations in the hsp70 5'flanking region (468-752 bp) had different effects on mRNA expression of HSP70, HSF1, Bcl-2 and Bax-$\alpha$. The ac genotypic cows showed higher expressions of HSP70mRNA, HSF1mRNA and Bcl-2mRNA/Bax-$\alpha$mRNA and lower ratio of apoptosis. These mutation sites can be used as molecular genetic markers to assist selection for anti-heat stress cows.
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제안 방법
Data on the abundances of HSP7QmRNA, HSF1mRNA and Bcl-2mRNA/Bax-amRNA were not independent, therefore, the transformation of the square root was used to normalize the data of the three groups in different genotypes to meet the requirements.
We do not know much about the expression of the genes mentioned above in the dairy cows undergoing heat stress. In order to explore the transmission molecular signals of bovine heat shock response, we designed four pairs of primers and analyzed both the ratio of apoptosis and the expression level of HSP70mRNA, HSF1mRNA and Bcl2mRNA/Bax-amRNA of the lymphocytes in the peripheral blood of dairy cattle at different daily-mean-temperatures.
We do not know much about the expression of the genes mentioned above in the dairy cows undergoing heat stress. In order to explore the transmission molecular signals of bovine heat shock response, we designed four pairs of primers and analyzed both the ratio of apoptosis and the expression level of HSP70mRNA, HSF1mRNA and Bcl2mRNA/Bax-amRNA of the lymphocytes in the peripheral blood of dairy cattle at different daily-mean-temperatures.
In this experiment, we chose the 5’-flanking region of the hsp70 gene to study.
In this experiment, we chose the 5’-flanking region of the hsp70 gene to study. PCR-SSCP assay showed that a mutation site and a CCAAT box existed in this region.
데이터처리
therefore, the transformation ofthe square root was used to normalize the data of the three groups in different genotypes to meet the requirements. To test variance of three groups among them, independent-samples F Test was conducted. Data shown are the mean±SD abundances of different genes.
성능/효과
(1993) through studying HSP70-2 mRNA expression in heat-shocked peripheral blood mononuclear cells from individuals with different hsp70-2 genotypes. Their data showed that PstI-homozygous individuals had slightly lower HSP70-2mRNA expression than heterozygous and PstI-homozygous individuals. Thus, interindividual and interracial differences in HSP70 expression could relate to regulatory mechanisms distinct from transcriptional regulation, as was also suggested by Lyashko et al.
There were no significant differences in HSP70-1mRNA accumulation and HSP expression between hsp70-1C and hsp70-1A homozygous cell lines. Their results thus suggested that the hsp70-1A and hsp70-1C bi-allelic promoter polymorphism did not affect HSP expression. Other authors reported a polymorphism in the coding region ofthe hsp70-2 gene and its association with diseases such as insulin-dependent diabetes mellitus (Pociot, 1993).
참고문헌 (22)
Ahn, S. G. and D. J. Thiele. 2003. Redox regulation of mammalian heat shock factor 1 is essential for Hsp gene activation and protection from stress. Gene and Development 17:516-528.
Aron, Y., M. Busson, B. S. Polla, D. Dusser, A. Lockhart and E. Swierczewski. 1999. Analysis of hsp70-1 gene polymorphism in allergic asthma. Allergy 54:165-170.
Badola, S., T. K. Bhattacharya and T. K. Biswas. 2004. A comparison on polymorphism of Bata-lactoglobulin gene in Bos Taurus and indicine ${\times}$ Taurine crossed cattle. Asian-Aust. J. Anim. Sci. 17:733-736.
Block, C. J., C. Chase and P. J. Hansen. 2002. Inheritance of resistance of bovinepreimplantation embryos to heat shock: Relative importance of thematernal versus paternal contribution. Mol. Repro. Dev. 63:32-37.
Chrenck, P., J. Huba and D. Vasicek. 2003. The relation between genetic polymorphism markers and milk yield in brown Swiss cattle imported to Slovakia. Asian-Aust. J. Anim. Sci. 16:1397-1401.
Deguchi, Y. and S. Kishimoto. 1990. Enhanced expression of the heat shock protein gene in peripheral blood mononuclear cells of patients with active systemic lupus erythematosus. Ann. Rheum. Dis. 49:893-895.
Dhillon, V. B., S. McCallum, P. Norton, B. M. Twomey, F. Erkeller-Yuksel and P. Lydyard. 1993. Differential heat shock protein overexpression and its clinical relevance in systemic lupus erythematosus. Ann. Rheum. Dis. 52:436-442.
Favatier, F., L. Bornman, L. E. Hightower, E. Gunther and B. S. Polla. 1997. Variation in hsp gene expression and Hsp polymorphism: do they contribute to differential disease susceptibility and stress tolerance? Cell Stress Chaperones 2:141-155.
Favatier, F., M. R. Jacquier-Sarlin, E. Swierczewski and B. S. Polla. 1999. Polymorphism in the regulatory sequence of the human hsp70-1 gene does not affect heat shock factor binding or heat shock protein synthesis. Cell. Mol. Life Sci. 56:701-708.
Holter, J. B., W. J. West, M. L. McGilliard and A. N. Pell. 1996. Predicting ad libitum dry matter intake and yields of Jersey cows. J. Dairy Sci. 79:912-921.
Holter, J. B., J. W. West and M. L. McGilliard. 1997. Predicting ad libitum dry matter intake and yield of Holstein cows. J. Dairy. Sci. 80:2188-2199.
Lyashko, V. N., V. K. Vikulova, V. G. Chernikov, V. I. Ivanov, K. A. Ulmasov and O. G. Zatsepina. 1994. Comparison of the heat shock response in ethnically and ecologically different human populations. Proc. Natl. Acad. Sci. USA 91:12492-12495.
Milner, C. M. and R. D. Campbell. 1990. Structure and expression of the three MHC-linked Hsp70 genes. Immunogenetics. 32:242-251.
Milner, C. M. and R. D. Campbell. 1992. Polymorphic analysis of three MHC-linked Hsp70 genes. Immunogenetics 36:357-362.
Mosser, D. D., A. W. Caron and L. Bourget. 1997. Role of the human heat shock protein HSP70 in protection against stressinduced apoptosis. Mol. Cell. Biol. 17:5317-5321.
Perfettini, J. L., R. T. Kroemer and G. Kroemer. 2004. Fatal liaisons of p53 with Bax and Bak. Nature Cell Biology. 6:386-388.
Pociot, F., K. S. Ronningen and J. Nerup. 1993. Polymorphic analysis of the human MHC-linked heat shock protein 70 (HSP70-2) and HSP70-Hom genes in insulin-dependent diabetes mellitus (IDDM). Scand. J. Immunol. 338: 491-495.
Ross, T., R. Olivier and L. Monney. 1998. Bcl-2 prolongs cell survival after Bax-induced release of cytochrome. Nature 391:496-499.
Shimizu, S., K. Nomura, M. Ujihara, K. Sakamoto, H. Shibata and T. Susuki. 1996. An allele-specific abnormal transcript of the heat shock protein 70 gene in patients with major depression. Biochem. Biophys. Res. Commun. 219:745-752.
Silvia, E., Valtorta and Miriam R. Gallardo. 2003. Evaporative cooling for Holstein dairy cows under grazing conditions. International Journal of Biometeorology 25:59-87.
Villunger, A., E. M. Michalak and M. Coultas. 2003. p53 and drug-induced apoptitic response mediated by BH3-only proteins puma and noxa. Science 302:1036-1038.
Yic, C., C. M. Knudson and S. J. Korsmeyer. 1997. Bax suppresses tumorigenesis and stimulates apoptosis in vivo. Nature 385:637-640.
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