Screening The Specific Binded Host-Proteins That Regulating Downstream Protein Of DNA5of Banana Bunch Top Virus

Banana bunch top disease (BBTD) is a one of main devastation disease for banana. In our Taiwan, Fujian, Guangdong, Guangxi, Hainan and Yunnan provinces, all of the banana regions almost infected with in different level of disease, serious harmed to banana which can be as high as80%, that has developed to devastating degree, and the impact for banana industry in China is also attention.Banana bunch top disease is caused by banana bunch top virus (BBTV). International Committee on Taxonomy of Viruses ruled that BBTV is the member of Nanoviridae family, Babuvirus. It consists of six compositions which about～1to～1.1kb in size. BBTV DNA1,3-6code Rep, CP, Mp, NSP and Rp connection protein, separately, but DNA2encods a unknown protein. At presently of studing abroad, we clear that BBTV DNA replicates and each protein of the function for the different isolate. While in the transcription regulation still blank in research. Therefore, the BBTV DNA components of un-coding areas regulation factors research is very necessary. This study uses new system view, that the un-coding areas on BBTV DNA5and the interaction of protein. we plan to screen some proteins that can combine with BBTV DNA5’s un-coding areas, that we can have a further knowledge on virus and pathogenesis mechanism.Foreign BBTV DNA copy and each group to the function of the protein un-coding have a more in-depth research in foreign. But what kind of contact between each protein and the host? And how to control the virus into the host body and cause a host of symptoms? such as lack of deliberate? Therefore, the role of the un-coding areas researching of BBTV DNA5component of regulation is very crucial. Banana bunch top virus DNA5codes a about20kD protein, the Rb-combined protein, the size is453code bases. Whether it has the function in regulating, and what kind of contact with host protein is the focus of this study.In this study, we use the yeast one hybrid method to study the interaction of BBTV DNA5un-coding area to the host. First, we use the banana bunch top virus Haikou isolate total DNA as a template, cloning the sequences of BBTV DAN5 un-coding areas, and at the ends are added enzyme cuting sites HindⅢ、Xhol Ⅰ,and adding the successfully constructed the carrier bait pABAi-5IR which can use in yeast one hybrid system. According to Clontech company yeast single hybrid kit, keeping the bait carrier restructuring to yeast cells, making the restructuring yeast cells growth in the defect of the type to SD medium. Second, we use the AbA antibiotics to test our bait carrier since the AbA antibiotic has a inhibition to the bait carrier,and the detection concentration for bait carrier is150ng/ml. Third, we use the banana bunch top virus Haikou isolate total RNA as a template to construct the cDNA library(library drops degrees at5.28X106), and co-transformation restructuring yeast cells. After a series of screening, we get one gene, binding proteins NAC.This research has obtained1host proteins which can interact with the BBTV DNA5IR--NAC binding protein. The NAC protein is belonged to one of the biggest protein family in plant transcription factors, that wide spread in terrestrial plants. Their interactions is very important in researching of the virus and the host interaction, but also it need continue to do BBTV other components (except DNA5) of banana regulation area and host protein interactions, and it makes a theoretical basis in banana resistance breeding.