Expression Analysis Of Receptor-Like Kinase Protein At Different Developmental Stages

Plant receptor-like kinase protein in the development of almost all the important metabolic processes related to known plant development, self-incompatibility, male sterility, resistance to disease and other life activities, stress resistance and play an important role in regulation. These kinases and their downstream proteins form a complex and orderly network, regulating the plant's normal growth and development and response to environment. So far, studies have been carried out functions of plant receptor-like kinase is a very limited number.We chose nine receptor protein kinase gene in rice, Oryza sativa through preparation for the receptor protein kinase antibody, to understand the rice protein kinase expression in different developmental stages of the information, for the function of accumulating data.1. we pass the epitope prediction, in vitro expression and purification of the protein kinase fragments. The purified protein used as immunogen to obtain a polyclonal antibody against the target gene. And to make full protein immunogen may be easier than access to sufficient quantities of high purity protein; with synthetic peptide immunogens compared to the cost of doing less. This experiment proved the feasibility of this route.2. Prokaryotic expression system, expressed five fragments of the protein kinase gene: Os04g49460, Os01g06280, Os03g21540, Os03g43670, Os01g48000, a large number of expressed and purified five proteins, polyclonal antibodies, fusion proteins used to detect antibodies that good immunogenicity.3. Of the four kinases genes (Os04g52860, Os03g12470, Os03g61310, Os07g02350) epitope fragment of the peptide synthesis, to synthetic peptides as antigen, prepared the appropriate polyclonal antibodies, antibody in different tissues of rice materials by Western blotting, to verify the specificity of antibody well, which further illustrates the preparation of polyclonal antibodies by the synthetic peptide line the feasibility of this technology.4. From a 93-11 different tissues tested materials for the Western blotting results, the prepared nine antibodies in rice protein has high specificity, get a clearer signal are basically the main belt. One Os04g49460 constitutive expression of a kinase protein, Os07g02350 and Os04g49460 the apparent molecular weight was consistent with the prediction, Os03g21540, Os04g52860, Os03g43670, Os03g12470, Os01g48000 and Os03g61310 protein of apparent molecular weight of less than the predicted molecular weight, Os01g06280 protein of apparent molecular weight higher than the predicted molecular weight, protein kinase Os03g21540 in the seedling stage is a period of significant expression of specificity.5. Posted on the NCBI library from 274 of more than 1 million EST analysis, nine kinase gene roughly uniform distribution of EST number, and Western blotting revealed the constitutive expression results consistent with the ATPase, HSP81- 3, EGF-1 alpha and RuBisCo, compared to the control gene, these EST kinase gene of the small number of shows that they all belong to low abundance of gene transcription.