Preparation Of Antibody Against OsRac1 And The Changes Of OsRac1 Expression In Rice Plants After RSV Infection

Rice stripe virus (RSV), a type member of Tenuivirus, distributes broadly in southern and northern rice producing areas of China and has caused great decreases to rice yield.G-protein is a kind of signal protein existing in almost all eukaryotes. OsRac1 is a homologue of human RAC in rice. Great attention has been paid to OsRac1 since it was cloned in the 1990s and results showed that it plays a central role in resistance to microbes.OsRac1 was successfully amplified by RT-PCR, and was cloned into pMD18-T vector, then into Prokaryotic expression vector pGEX-4T-3 after being cleaved. The vector was transformed into Escherichia, coli DH5a. Positive recombinants expression plasmids were screened. The sequence was determined to ensure the success of gene cloning and absence of frameshift mutations. The results indicated that the Molecular Weight of the inserted sequence is about 23 kD.Recombinant expression plasmids were transformed into E.coli BL21 (DE3) and were induced by IPTG, results of the SDS-polyacrylamide gel electrophoresis were consistent with the expected size of the fusion protein band. The antiserum against OsRac1 was prepared. Western blot results showed that the prepared antiserum and the induced fusion protein had specific reaction.At present, only qualitative research on OsRac1 is available. To test the possible involvement of OsRac1 in rice-RSV interaction, Western blot experiments using the polyclonal antibody raised against OsRac1 were carried out. The results showed that expression of OsRac1 protein was more abundant in the healthy rice plants than that in the RSV-infected rice plants.