Analysis On One Novel Protein OsHSPBP Which Interacted With OsHSP90

Rice (Oryza sativa L.) is one of the main crops in our country even in the world. Abiotic stress damage is one of the important factors that make its production potential could not be fully exploited. Therefore, the research of adverse resistance is always the important task and finding resistant gene has been the hotspot on both the studying of rice genetic resources and variety improvement researches. In recent years, along with the development of the theories and technology of the molecular biology, the research of stress genes has made great progress. Some genes coding proteins related to stress have been isolated and transferred to the rice, thereby, the transgenic plants with high stress resistance have been acquired, which is helpful to comprehend the adaptive mechanism of rice under adversity and to guid to filter and cultivate the rice with resistance.Heat Shock Protein(HSP) exists in the biological universe ubiquitously. HSP not only can perform effectively under stress condition, but also exists in the cells on normal state participating in some important cell activities. The biological function of HSP is diversified so that it can improve the tolance ability of cell against adversity and it also can be molecular chaperone. The current research has shown that HSP is related to the plants' growth,development, differentiation, cell signal transduction and phenotypic variation. There are differences among the various HSP in sequence and sizes. In order to study the genetic structure, expression characteristics and the function of OsHSP90 systematically, this research 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 OsHSPPBP. Results are as follows:1.According to the approaches provided by Clontech and making use of the yeast two-hybrid tecnology by taking OsHSP90 as bait protein, the unknown function protein is filtered from the low temperature and drought cDNA library of rice by using the PEG/LiAc method to transform yeast cell.2.Analyzing OsHSPBP by comparing in the data-bases, follows could be find: cDNA of this gene is 1321 bp long, its coding area lies in 184bp-918bp encoding 244 amino acid residues. molecular weight is predicted to be about 25.5 KD and isoelectric point is about 9.49 . Comparing the amino acid homology, OsHSPBP is found high similarity in the protein to other plants, and the similarity in the protein to arabidopsis thaliana(AAG48784) , corn (ACG33139) and grape (CAN83805) are respectively 32.46%, 70.40% and 30.66% and the similarity between the functional areas from tify to CCT₂ are obvious higher than that in other areas. In addition, phylogenetic tree analysis shows that sequence similarity of the protein in the rice OsHSPBP and corn is higher than that in arabidopsis thaliana and grape.3.Using the method of semi-quantitative RT-PCR and real-time quantitative PCR to analyze the tissues expression of OsHSPBP and OsHSP90 in the conditions stressed by low temperature, high temperature, drought and high salt in different physiological stages. Both of them are found to have different expressions in degrees before dealing with adversity stress, which are consistent in the conditions of heat-shock, low temperature and drought and express as tissue specific expressed genes in internodes, nodes, sheaths ,branches and glumes in different physiological stages.4. Constructing expression vector of sense and antisense cDNA fragment of OsHSPBP genes and establishing the genetic transform system for rice mediated by agrobacterium tumefaciens: direct sequence transfer to the rice for over expression is to amplify the physiological function of the gene; reverse sequence transfer is to prevent the expression of OsHSPBP in the rice by homologous sequence inhibition, consequently, to study the physiological function of the gene expression in resistant response of the rice. Meanwhile, to filter the seedings with transfection by using the method of antibiotics is on study now.