The S1 Gene Eukaryotic Expression Vector Construction And The Agrobacterium-mediated Transformation Of The Preliminary Study On Rice

Viral disease which influences the output and quality of rice is a serios problem in provisionment.Tranditional approaches like crop-dusting to defend this desease shall make environmental-polluting.At present,cultivating virus resistant rice by genetic engineering is one of the methond to solve this contradiction.The rice stripe disease is a kind of rice-damaging entomophilous disease caused by RSV.In the worldwide and China,the disease occurred devastatingly and repeatedly.Two subspecies of Cultivars were found resistant on RSV,which are the variety of O.sativa var.Japonica,weak in disease resistance but superior in quality,and the variety of O.sativa var.Indica,strongly disease-resistant but poor in quality.The research showed that the crude extract of Indica include the factor of bioactive resistance which,after the salting out,hydrophobic,and ion exchange chromatographic tests,could still highly restrain the composition of RSV.The research mainly focuses on the a protein peptide which is related to duplication of RSV resistance has,was separated from the RSV-resistant Indica.In analyzing the end of this protein peptide,by the means of PCR and synthetic method the full length cDNA of the gene was gained and named as s1.In the research and analysis of the genomic DNA of sl,it was found that sl sequese an unknown protein and has eight introns,and Chloroplasts in its sequence.In order to further study the coding function of the s1 protein,constructed a Yeast Expression Plasmids pYES2/CT,and the expression protein was obtained.The s1 was also cloned into plant expression vector pCAMBIA-1302 and then the recombinant vector was transformed into Agrobacterium strain LBA4404.The s1 gene was transformed into rice(Oryza stiva.L) by the Agrobacterium tumefaciens in order to improving disease resistance to viral disease.Many transgenic calli and regenerations were obtained and screened by PCR.Southern bloting and northern bloting will be used to test these transformations in the future.This experiment also used onion epidermal cells for the s1 gene initial subcellular localization,and found that the gene location in the nucleus.