中文摘要：

香蕉束顶病毒DNA2组分编码蛋白的功能尚不清楚，为了揭示其编码蛋白的功能，以本实验室保存的BBTV Haikou4分离物总DNA为模板进行PCR扩增。利用Gateway重组技术将BBTVDNA2ORF构建到酵母双杂交系统（ProQuest^TM Two—Hybrid System，Invitrogen）的诱饵载体pDEST-32中，并通过酵母表型验证、诱饵质粒毒性验证以及自激活验证。结果显示．转有pDEST32-DNA2 ORF质粒的MaV203酵母菌具有弱的自激活背景，在3-AT浓度为25mmol／L的SeC—Leu—Trp—His平板上生长较弱。但在3一AT浓度为50mmol／LSeC—Leu—Trp—His平板上不生长，表明诱饵质粒背景自激活可被浓度为50mmol／L的3-AT所抑制。在该条件下共转pDEST32-DNA2ORF和文库质粒到MaV203感受态细胞进行互作蛋白筛选，通过PCR和测序等手段最终鉴定35个候选蛋白基因，分别编码转录调控因子、生长发育相关蛋白、代谢相关蛋白、抗逆相关蛋白和未知蛋白等。酵母双杂交BBTVDNA2ORF诱饵质粒的构建及其与寄主互作蛋白的筛选，为揭示DNA2组分的蛋白功能提供了科学线索。

英文摘要：

The function of the protein encoded by Banana bunchy top virus （BBTV） DNA2 is still unknown. In order to reveal the function of this protein, DNA20RF was cloned from BBTV Haikou4 total DNA and gateway recombination was used to build the yeast bait vector pDEST32-DNA20RF via yeast two-hybrid system （ProQuestTM Two-Hybrid System, Invitrogen）. The pDEST32-DNA20RF bait vector can be used for protein- protein interaction screening after test by yeast phenotypic verification, bait plasmid toxic verification and self-activation verification. The results showed that pDEST32-DNA20RF/MaV203 weakly grew on SeC-Leu-Trp- His YPAD plate with 25 mmol/L 3-AT, but no colony showed up on SeC-Leu-Trp-His YPAD plate with 50 mmol/ L 3-AT. In this condition, the pDEST32-DNA20RF and AD-Library plasmids were co-transferred into MaV203 competent cell to screen the interaction proteins. The results showed that 35 candidate protein genes were obtained, encoding transcription factors proteins, growth-associated proteins, metabolism-associated proteins, resistance associated proteins and unknown proteins. The study of BBTV DNA20RF bait plasmid construction and interacting proteins screening could facilitate the identification of the functions of BBTV DNA2 encoded protein.