大针茅质膜水孔蛋白亚家族pip基因cdna序列的克隆及分析-生物化学与分子生物学专业论文.pdf.docxVIP

目录1引言................................................................11.1大针茅植物简介..................................................11.1.1大针茅植物特征................................................11.1.2大针茅植物的研究概况..........................................11.2水孔蛋白简介....................................................21.2.1水孔蛋白的发现................................................21.2.2水孔蛋白的分类与多样性........................................21.2.3水孔蛋白的结构特征............................................31.3研究目的与意义..................................................42材料与方法..........................................................42.1实验材料........................................................42.2仪器和试剂......................................................52.3总RNA提取52.4特异性引物的设计................................................6SgPIP中间cDNA的引物6SgPIP3′RACE的特异性引物6SgPIP5′RACE的特异性引物6SgPIP全长cDNA扩增的引物72.5反转录PCR反应7SgPIP中间cDNA的克隆83′RACE技术克隆SgPIP的3′末端cDNA 9SgPIP3′末端cDNA第一链的合成9SgPIP3′末端cDNA的嵌套PCR反应95′RACE技术克隆SgPIP的5′末端cDNA 11SgPIP5′末端cDNA第一链的合成11SgPIP5′末端cDNA的嵌套PCR 11SgPIP全长cDNA的克隆122.10生物信息学分析.................................................123结果与分析.........................................................133.1总RNA的提取及检测13SgPIP中间cDNA的克隆及分析14SgPIP中间cDNA的PCR扩增产物检测14SgPIP中间cDNA的阳性克隆鉴定143.3SgPIP3′末端cDNA的克隆及分析15SgPIP3′末端cDNA的PCR扩增产物检测15SgPIP的3′RACE阳性克隆鉴定16SgPIP5′末端cDNA克隆及分析16SgPIP的5′末端cDNA的PCR扩增产物检测16SgPIP的5′RACE阳性克隆鉴定18SgPIP全长cDNA的克隆及分析18SgPIP全长cDNA的PCR扩增产物检测18SgPIP全长cDNA的阳性克隆鉴定193.6SgPIP的生物信息学分析223.6.1SgPIP的核苷酸与氨基酸序列分析及命名223.6.2SgPIP蛋白疏水/亲水性的预测和分析263.6.3SgPIP蛋白跨膜结构域的预测和分析273.6.4SgPIP蛋白三级结构的预测294讨论...............................................................304.1大针茅SgPIP的中间cDNA的克隆304.2大针茅SgPIP的3′和5′末端cDNA克隆305结论...............................................................31致谢..............................................................32参考文献.......................................................33作者简介.......................................................371图1水孔蛋白模拟结构“水漏模型”..............................