The Constructions Of Over-expressed,Anti-sense And RNAi Expressing Vectors And Their Genetic Transformation To Rice (Oryza Sativa, L.)

RACK1, Receptor for Activated C Kinase1, is a scaffold protein and plays important roles in intracellular responses. Several results have showed that RACK1 also existed in plants. In Arabidopsis thaliana, three RACK1 genes have been cloned and have been involved in the response to hormones and drought stress. Using AtRACK1 as a template, We found two genes in rice genome highly homologous to that encoding AtRACK1. In the present study, we tried to clone OsRACK1 gene from stems and leaves of Oryza sativa subsp. japonica cv. Nipponbare using RT-PCR and to construct different expressing vectors (i.e., Over-expressing- and suppressing- (using antisense and RNA interference, RNAi) vectors). And then, we transformed rice callus using agrobacterium tumefaciens system. The results showed that ,1. OsRACK1 has been cloned from the stems and leaves of Oryza sativa subsp. japonica cv. Nipponbare. Gel-electrophoresis and DNA sequencing analysis showed that the size of the cloned OsRACK1 was 1199 bp, which contained the whole edcoding sequence of the gene.2. We successfully constructed three kinds of expression vectors, that is over-expressing vector which was drived by 35S promoter, anti-sensed and RNA interferred OsRACK1 vectors,and successfully transferred these three kinds of expressing vectors to rice callus using agrobacterium tumefaciens approach, and have screened three kinds of transformed rice plants. Histochemical analysis of reporter gene GUS gene and PCR assasy showed that target gene have been transferred into rice cells.