Phenotypic Identification And Function Exploration Of Receptor-like Protein Kinase OsRLK381

In the growth process, the plant can accept external signaling molecules and the various signals between cells in the body, which through a series of signal transduction pathways showed the physiological and biochemical reactions. The first plant receptor-like protein kinase (RLKs), ZMPK1genes successfully cloned, opened the prelude of the researchers on the study of plant RLKs. After that, researchers found a large number of genes coding and separating of RLKs.At present,about600RLKs have been found in arabidopsis thaliana.That is2.15%of the number of genes encoding proteins in that plant, and about1130in rice. Although the plant has the huge number of receptor protein kinases, its function, signal transmission ways and the ligand recognition is still poorly understood by now. some bioinformatics analysis,dominant negative control,rice plant phenotype identification and gene transformation function attempt conducted in this paper of rice protein kinase receptor OsRLK381(coding genetic Os02g19550).And we get the following main research results:(1) By the use of bioinformatics analysis, the nucleic acids and amino acids OsRLK381coding sequence have been analyzed.We found that OsRLK381is a membrane protein, which have a transmembrane domain structure, a L-lectin extra cellular structure domain and a tyrosine kinase domain inside the cell structure. Evolution analysis shows that this gene is in an independent evolution branch(2) We took kitaake wild-type rice as experimental material, rice total RNA as template, and amplified OsRLK381gene by RT-PCR, which was about1776bp,encoded591amino acids,as expected.(3) We successfully constructed Dominant negative control expression vector pCAMBI1301-OsRLK381, and confirmed by sequencing and PCR. Then the stable transgenic lines were generated through rice-mediated transformation. Through screening with hygromycin, western-blot and RT-PCR,we got three genetically modified strains OsRLK381-1OsRLK381-12、OsRLK381-17. Studies shown that the transgenic plants appear a phenotype of tall and with long spikes when Compared with wild type. We speculate that OsRLK381may affect internode elongation at vegetative stage. It may also affect the development Ear primordia at early of Reproductive growth.(4)By constructing prokaryotic expression vector pCOLD-OsRLK381R,and tra n-sforming the vector into E.coli BL(21)star, we obtained the engineering bacteria of prokaryotic expression-BL(21)star-pCOLD-OsRLK38lR. By using IPTG to induc e OsRLK381R gene and western-blot technology.the Fusion protein have been succ ess-ful expressed. The Molecular weight is75KD.