摘要
水是生命的重要要素,水的流入和流出细胞主要是通过细胞膜上存在的水孔蛋白完成的。植物水孔蛋白可分为四个家族,其中PIP1作为一种质膜内在蛋白(Plasma membrane intrinsic proteins, PIPs)在植物的生理生化过程中具有重要作用。
本课题通过构建碱茅的cDNA文库,并通过对文库库容进行检测,得到了一个具备较高的库容,并且具有完整性和代表性的碱茅cDNA文库。从碱茅的cDNA文库中克隆得到了水孔蛋白基因(PutPIPl),对基因的生物学信息、表达特性、酵母亚细胞定位以及转基因植株的逆境适应性进行解析。通过对PutPIP1基因的生物学信息分析,表明PutPIP1基因属于水孔蛋白家族中的质膜内在蛋白的PIP1亚类,而且具有6个跨膜结构。通过对PutPIP1基因的表达特性的分析,PutPIP1在野生碱茅各组织器官的转录水平特性进行研究,表明PutPIP1在碱茅的各部位中均有一定量的表达,并且该基因的表达具有一定范围内的组织特异性。将重组质粒pYES2-GFP-PutPIP1转入酵母细胞,PutPIP1与绿色荧光蛋白融合,同时以GFP为对照,诱导该基因表达,证明PutPIP1定位于酵母的质膜上。在逆境条件下,PutPIP1基因的表达在一定范围内增强酵母对NaCl和Na2CO3的耐受性,在低NaHCO2逆境条件下,没有表现出耐性趋势。
This topic from the alkali Lemongrass cDNA cloning got the water hole protein gene, on gene expression of biological information, features, and yeast subcellular localization of transgenic plants modified for resolution.
Through constructing cDNA library of Puccinellia,and through the library storage capacity to detect, we get a higher storage capacity cDNA library Puccinellia, which has integrity and representative.Through the PutPIP1 gene analysis of biological information, suggesting that the PutPIP1 gene belongs to the Aquaporin family inherent in the plasma membrane protein PIP1 subdivisions, and has six transmembrane structure.Through the PutPIP1 gene expression in characteristics of PutPIP1 in the wild, chinampoensis organizations organs of transcription characteristics to undertake research, indicating that PutPIP1 in Puccinellia in all parts of the expression in a certain amount,and organized specific gene expression in a certain range.
Recombinant plasmids will turn into yeast cells, pYES2-GFP-PutPIP1 and green fluorescent protein mingle with GFP as control and guide the PutPIPl gene expression to prove that the yeast is oriented to the plasma membrane.Under certain conditions, the PutPIP1 gene expression show a enhancing tolerance in Na2CO3 and NaCl, and without enhanced NaHCO3 resistance of salt.
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