Expression Of Related Genes That Resist Rice Strip Virus In E.coli

The rice stripe disease is a kind of rice-damaging entomophilous disease caused by Rice strip virus(RSV).In Japan,Korea 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.Our research mainly focuses on the related genes that are of rice stripe disease resistance.Now the full length cDNA of the gene was gained and named as sl.In order to further study the coding function of the S1 protein,a Prokaryotic Expression Plasmids(pET30a) was constructed respectively in S1 protein and the expression protein was obtained.The result of SDS-PAGE showed that successful expression of a 32kD fusion protein was achieved by induction.We set up gradient of times to improve the condition of Prokaryotic expression.We found that the expression of six hours to reach the highest.Temperature in the induction,we found that 37℃was easy to format inclusion bodies.To reduce the emergence of inclusion,we lowered the temperature to 28℃.Finally we got soluble proteins.And furthermore it was indicated by immunoblotting that the fusion protein was expressed correctly.We choose the method of Pull-Down to study the target protein's role.In order to find protein interaction,we made s1 protein as "bait" and rise-wide protein as "prey".The RSV-resistant protein was successfully obtained by prokaryotic expression system.We found that there are protein in rice can interact with the target protein.So the specific role of the target protein was available.