小麦硫转运蛋白基因rt-pcr半定量检测体系的建立.rar
小麦硫转运蛋白基因rt-pcr半定量检测体系的建立,小麦硫转运蛋白基因rt-pcr半定量检测体系的建立14页 1.3万余字，包含完整毕业设计及答辩ppt[摘要] 应用rt-pcr技术， 以tubulin为内参基因，小麦硫转运蛋白基因(sul)为检测基因，优化mg2+浓度、退火温度、循环次数、引物浓度和taq酶浓度，获得适宜的pcr反应参数，建立一个相对稳定、准确、特异的...
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小麦硫转运蛋白基因RT-PCR半定量检测体系的建立
14页 1.3万余字，包含完整毕业设计及答辩ppt
[摘要] 应用RT-PCR技术， 以Tubulin为内参基因，小麦硫转运蛋白基因(Sul)为检测基因，优化Mg2+浓度、退火温度、循环次数、引物浓度和Taq酶浓度，获得适宜的PCR反应参数，建立一个相对稳定、准确、特异的检测Sul表达量的半定量体系，为进一步研究小麦硫转运蛋白表达调节机制奠定技术基础。
[关键词] 小麦；Tubulin；硫转运蛋白基因；RT-PCR；半定量PCR
To Establish a Method of Semiquantitative Polymerase Chain Reaction for Detecting Sul Gene Expression
Abstract To establish a method of semiquantitative polymerase chain reaction for detecting Sulphate transporter gene expression in wheat roots，we established a semiquantitative PCR by putting primer of Tubulin served as internal reference gene and primer of target gene in the same conditions and optimizing experiment parameters for PCR such as the concentration of Mg2+ and cycle times etc. This method is usable to test the expressive level of Sul gene in the wheat roots.
Key words：wheat；Sulphate transporter；RT-PCR；semiquantitative PCR
参考文献
[1] C. W．迪芬巴赫，G.S．德维克斯勒，主编．(卢柏松、赵东，译) PCR技术实验指南．北京：科学出版社，第2版．1998．10—15．
[2] J.萨姆布鲁克 E.F弗里奇 T.曼尼阿蒂斯 分子克隆实验指南第3版[M]．科学出版社
[3] 张捷，定量和半定量PCR技术及其临床意义，北京医科大学学报，vol.31 No.5 Oct.1999
[4] 李珊珊等，3种mRNA检测方法比较，Journal of Henan Medical University Mar.2000 Vol.35 No.2
[5] 董爱香等，小麦春化相关特异性PCR扩增体系的建立，山西农业大学学报 A 1000 162X（2000）03-0205-03
[6] 黄春等，RT-PCR半定量法检测人脐静脉内皮细胞t-PA和PAI-1mRNA表达，福建医科大学学报1999年第33卷第3期黄春等
[7] 张继澍主编，植物生理学，世界图书出版公司
[8] 沈同，王镜严主编，生物化学，高等教育出版社，第三版
[9] Leustek T and Saito K (1999) Sulfate transport and assimilation in plants. Plant Physiol 120: 637-643）
[10] Dreyer I.,Horeau C.,Lemaillet G., Zimmermann S., Bush D.R., Navarro A.R., Schachtman D.P., Spalding E.P., Sentenac H.,Gaber R.F. Identification and characterization of plant transporters using heterologous expression systems J. Exp. Bot. Vol.50, Special Issue,pp.1073-1087,June 1999
[11] Hawkesford MJ and Smith FW (1997) Molecular biology of higher plant sulphate transporters. In Sulphur Metabolism in Higher Plants (ed. Cram WJ, De Kok, L J, Stulen, I, Brunold, C, and Rennenberg, H.) Backhuys Publishers, Leiden, The Netherlands pp 13-25.
[12] Smith, F.W., Rae, A.L. and Hawkesford, M.J. (2000) Molecular mechanisms of phosphate and sulphate transport in plants. Biochim. Biophys. Acta 1465: 236-245.
[13] Clarkson, D.T., Hawkesford, M.J., Davidian, J-C. and Grignon, C. (1992) Contrasting responses of sulphate and phosphate transporters in barley (Hordeum vulgare L.) roots to protein modifying reagents and inhibition of protein synthesis. Planta 187, 306-314
[14] Hawkesford MJ and Smith FW (1997) Molecular biology of higher plant sulphate transporters. In Sulphur Metabolism in Higher Plants (ed. Cram WJ, De Kok, L J, Stulen, I, Brunold, C, and Rennenberg, H.) Backhuys Publishers, Leiden, The Netherlands pp 13-25.
[15] Takahashi, N. Sasakura, M. Noji, K. Saito, Isolation and characterization of a cDNA encoding a sulfate transporter from Arabidopsis thaliana FEBS Lett.392 (1996) 95^99.
[16] H. Takahashi, M. Yamazaki, N. Sasakura, A. Watanabe, T. Leustek, J. de Almeida Engler, G. Engler, M. Van Montagu, K. Saito, Regulation of sulfur assimilation in higher plants: a sulfate transporter induced in sulfate-starved roots plays a central role in Arabidopsis thaliana. Proc. Natl. Acad. Sci. USA 94 (1997) 11102^11107.
14页 1.3万余字，包含完整毕业设计及答辩ppt
[摘要] 应用RT-PCR技术， 以Tubulin为内参基因，小麦硫转运蛋白基因(Sul)为检测基因，优化Mg2+浓度、退火温度、循环次数、引物浓度和Taq酶浓度，获得适宜的PCR反应参数，建立一个相对稳定、准确、特异的检测Sul表达量的半定量体系，为进一步研究小麦硫转运蛋白表达调节机制奠定技术基础。
[关键词] 小麦；Tubulin；硫转运蛋白基因；RT-PCR；半定量PCR
To Establish a Method of Semiquantitative Polymerase Chain Reaction for Detecting Sul Gene Expression
Abstract To establish a method of semiquantitative polymerase chain reaction for detecting Sulphate transporter gene expression in wheat roots，we established a semiquantitative PCR by putting primer of Tubulin served as internal reference gene and primer of target gene in the same conditions and optimizing experiment parameters for PCR such as the concentration of Mg2+ and cycle times etc. This method is usable to test the expressive level of Sul gene in the wheat roots.
Key words：wheat；Sulphate transporter；RT-PCR；semiquantitative PCR
参考文献
[1] C. W．迪芬巴赫，G.S．德维克斯勒，主编．(卢柏松、赵东，译) PCR技术实验指南．北京：科学出版社，第2版．1998．10—15．
[2] J.萨姆布鲁克 E.F弗里奇 T.曼尼阿蒂斯 分子克隆实验指南第3版[M]．科学出版社
[3] 张捷，定量和半定量PCR技术及其临床意义，北京医科大学学报，vol.31 No.5 Oct.1999
[4] 李珊珊等，3种mRNA检测方法比较，Journal of Henan Medical University Mar.2000 Vol.35 No.2
[5] 董爱香等，小麦春化相关特异性PCR扩增体系的建立，山西农业大学学报 A 1000 162X（2000）03-0205-03
[6] 黄春等，RT-PCR半定量法检测人脐静脉内皮细胞t-PA和PAI-1mRNA表达，福建医科大学学报1999年第33卷第3期黄春等
[7] 张继澍主编，植物生理学，世界图书出版公司
[8] 沈同，王镜严主编，生物化学，高等教育出版社，第三版
[9] Leustek T and Saito K (1999) Sulfate transport and assimilation in plants. Plant Physiol 120: 637-643）
[10] Dreyer I.,Horeau C.,Lemaillet G., Zimmermann S., Bush D.R., Navarro A.R., Schachtman D.P., Spalding E.P., Sentenac H.,Gaber R.F. Identification and characterization of plant transporters using heterologous expression systems J. Exp. Bot. Vol.50, Special Issue,pp.1073-1087,June 1999
[11] Hawkesford MJ and Smith FW (1997) Molecular biology of higher plant sulphate transporters. In Sulphur Metabolism in Higher Plants (ed. Cram WJ, De Kok, L J, Stulen, I, Brunold, C, and Rennenberg, H.) Backhuys Publishers, Leiden, The Netherlands pp 13-25.
[12] Smith, F.W., Rae, A.L. and Hawkesford, M.J. (2000) Molecular mechanisms of phosphate and sulphate transport in plants. Biochim. Biophys. Acta 1465: 236-245.
[13] Clarkson, D.T., Hawkesford, M.J., Davidian, J-C. and Grignon, C. (1992) Contrasting responses of sulphate and phosphate transporters in barley (Hordeum vulgare L.) roots to protein modifying reagents and inhibition of protein synthesis. Planta 187, 306-314
[14] Hawkesford MJ and Smith FW (1997) Molecular biology of higher plant sulphate transporters. In Sulphur Metabolism in Higher Plants (ed. Cram WJ, De Kok, L J, Stulen, I, Brunold, C, and Rennenberg, H.) Backhuys Publishers, Leiden, The Netherlands pp 13-25.
[15] Takahashi, N. Sasakura, M. Noji, K. Saito, Isolation and characterization of a cDNA encoding a sulfate transporter from Arabidopsis thaliana FEBS Lett.392 (1996) 95^99.
[16] H. Takahashi, M. Yamazaki, N. Sasakura, A. Watanabe, T. Leustek, J. de Almeida Engler, G. Engler, M. Van Montagu, K. Saito, Regulation of sulfur assimilation in higher plants: a sulfate transporter induced in sulfate-starved roots plays a central role in Arabidopsis thaliana. Proc. Natl. Acad. Sci. USA 94 (1997) 11102^11107.
