Analysis On The Function Of One Novel Gene 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. With the development of plant molecular biology, 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, this has opened up a new era for corp breeding.Plant is often subjected to biotic stress and abiotic stress under growth and development,the researchers have been focus on stress signal transduction.Under stress conditions, plant itself can perceive and conduct stress signal, start lifting stress-related gene expression, and then activate the appropriate protection of metabolic pathways.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. HSP90 family is a kind of molecular weight about 90 kD heat shock protein, it has helped protein folding, maintained the integrity of protein structure, and regulated the expression of cytoplasmic proteins. 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 was 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 found:cDNA of this gene is 1321bp long, its ORF 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 maize (ACG33139) similarity is the highest 70.40% and the similarity between the functional areas from tify to CCT₂ are obvious higher than that in other areas.3. Using the method of semi-quantitative RT-PCR and real-time quantitative PCR to analyze the tissues expression of OsHSPBP in the conditions stressed by low temperature, high temperature, drought and high salt in different physiological stages. It is found to have different expressions in degrees before dealing with adversity stress:the expression is first increased and then reduced under high temperature and high salt; under low temperature and drought, the expression is increased. Meanwhile, it is not 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 to study the physiological function of the gene expression in resistant response of the rice. At the same time, OsHSPBP-GFP is built into the 1301 vector for subcellular localization of the OsHSPBP.