Study On The Founction Of RDV S6,S11

RDV is made up of twelve pieces of ds RNA, among them, six genes are to encode structural protein, others are to encode nonstructural protein. S6 and Sll belong to the latter. My experiment has primarily verified that Pns6 is motive protein; Pnsll is nuclear acid binding protein. I have primarily identified its nuclear acid binding domain, fatherly, it can go into chloroplast by some way. The study on RDV motive protein Pns6 is based on complementary experiment of Pns6 and motive disfigurement plant. The procedure is as follows: with the invective carrier of pvx motive disfigurement the mutation clones of PVX cDNA and CaMV 35S -RDV S6 with GUS and GFP gene respectively are transformed into tobacco epidermis by gene gun bormbardment or cotransformation to express protein between cells. The results testified that the protein encoded by S6 can complete the motive function between cells of pvx mutation, indicated that Pns6 is motive protein between RDV cells. So, this kind of virus' motive protein between cells has been found and verified firstly and internationally. In the same time, I have studied the motive protein between cells encoded by S6. The fusion gene of S6 and EGFP has been cloned into plant expression carrier and expressed protein in the N.tabacum. I found that the protein encoded by S6 was expressed specially both in thread between cells and nucleoluses. The expression in cytoplasm is unrelated to protein made of cell framework. The assay from the hyperploid's thin slices of healthy rice and infected ones and immunocolliod tagged with gold of protein encoded by S6 and differential antibody of outer capsid, indicated that the protein encoded by S6 specially orientates in the thread between cells of the rice infected by RNA and the viral duplex. The result is consistent with the result marked by GFP. Besides, from the study above, I also found that there was not viral particle analog beside the thread between cells, which suggested that RDV moving through the thread between cells perhaps is not in the form of viral particles. Due to having not suffice learning about Pns6 and Pns11. Furthermore, they are both nonstructral protein, their content in the infected rice and leafhopper is very low, and is difficult to be purified. Now, the best way is to express the RDV protein largely through the viral system of nucleolus or nonnuclelous to supply for study. In this thesis, Pns6 and Pnsll have been expressed through the expression system ofpichia.pastoris, then, they were verified by Western blotting. S6 and S11 have also been studied through Yeast two-hybrid system, in this study, four genes that can interact with S6 have been found, and five genes that can interact with S11 have also been found. Then, they have been sequenced: most of them are 3' end DNE sequence. I gained the whole length DNA fragments through 5' RACE. Following, 1 compared the gene sequence through BLASTEN searching in order to found the highly homologous gene that can be used as a model for further study. This thesis has studied the related genes in molecular and cell standard and enhanced the learning about Pns6 and Pnsll, which has important academic basement on RDV molecular virology and the interaction of RDV and host plants.