Analysis Of A New Unkown Protein Gene Which Interact With OsHsp90

The plant is a open system,living in the natural environment, whose growth and development are affected by the environment. In the natural environment,the environment which is not fit for their growth and development appears unavoidably, it affects their normal development.In Recent years, due to the deterioration of the global environment, the bad wheather such as low temperature, high temperature, drought and so on affect the normal growth and development of the plant seriously,especially the crop. The rice(Oryza sativa L.)as one of the most important grain crops in our courntry ranks first among various grain crops on the total area, the total output and the yield per unit. But recently, more and more worse environment affect the output of the rice seriously, therefore, the research on the tolerance of rice is always the one of the most important issues of rice research, the discovery of the gene for stress tolerance have been a hotspot on the research of rice genetic resourses and breed improvement, and they have found many genes for stress tolerance,one typical example of which is heat-shock protein.Heat-shock protein is a kind of ubiquitous protein in biosphere, which can express efficiently under stress and involve in some kinds of cellular physiological activity. The bio-function of Hsp has the characteristic of diversity, except for increasing the tolerence under stress and acting as a molecular chaperone, relating to the normal growth of the plant, development and differentiation, signal transduction in cell, phenotypic variation and so on. This study takes OsHsp90 as bait protein to filter its interact protein by using yeast two-hybrid system,then analyzes the function of the new gene of Os-unkown protein (OsUnP), in order to further analyzing and showing the function of the rice heat-shock protein family and their binding protein on the interaction and signal transduction and so on, laying a solid foundation of researching the machanism of the interaction between OsHsp90 and their binding protein. Results as following:1. According to the approaches provides by Clintech and making use of the yeast two-hybrid tecnology by taking OsHsp90 as bait protein, the unknown function protein is filtered fromthe low temperature and drought cDNA library of rice.2. Analyzing OsUnP by comparing in the data-bases, following could be found:cDNA of this gene is 1058bp long, its coding area lies in 138bp-666bp encoding 176 amino acid residues.molecular weight is 19.7KD. Comparing the animo acid homology, OsUnP is found high similarity in the protein sequence to other plants:the similarity in the protein to Oryza sativa subsp.Japonica (Q656U1), Oryza sativa subsp.Indica(A2WRH6), Zea mays(B6T6A1) and Physcomitrella patens subsp.Patens(A9SC52) are respectively 99%, 98%,73% and 48%. But to the exogenous contrast-Glycine max(C6SZA4), which belongs to the H3 family, is only 11%. Furthermore, because of the existing of DUF866 domain, they may have the same or the similar function.3. Using the method of semi-quantitative RT-PCR to analyzing the tissues expression of OsHsp90 and OsUnP under the stresses such as low temperature, high temperature, drought and high salinity and so on, and in different physiological stage. Both of them are found to have different expressions in degrees before dealing with adversity stress, which show consistent trend under high temperature, high salinity, and opposite trend under drought; they express in internodes, nodes, sheaths, branches and glumes in different physiological stages.4. Constructing expresing vector of sense, antisense and GFP cDNA fragment of OsUnP and establishing the genetic transform system for rice mediatied by agrobacterium tumefaciens to importing to the rice callus, then forming the transgene plants. Through researching the morphologic and physiologic indexes' change of the transgene plants in different physiological stages and under stresses, showing the function of this gene under stresses, laying the foundation of the application of this gene in the breeding for stress tolerance.