Screening Interaction Proteins of Rubber Elongation Factor (REF) by Yeast Two-hybrid System
应用酵母双杂交系统筛选橡胶延长因子REF的互作蛋白

The interaction proteins of rubber elongation factor (REF) were screened using yeast two-hybrid system. A bait vector pBD-GAL4-REF without auto-activation was constructed by subcloning ORF cDNA of REF gene into the yeast two-hybrid vector pBD-GAL4, and the pAD-GAL4-2.1 cDNA prey library was prepared with mRNA extracted from the latex of rubber tree (Hevea brasiliensis). The bait and prey vectors were then co-transformed into YRG-2 yeast competent cells by two-step method. Five proteins interacted with REF were identified by yeast two-hybrid screening method, including two of REF family proteins, and small rubber particle protein (SRPP), translation control of tumor protein (TCTP), elicitor-responsive protein and ubiquitin-conjugating enzyme E2. The data suggested that REF could not only interact with high homologous proteins, but also with other proteins, such as TCTP, and E2, etc. These protein-protein interactions might help to revealing the biological function of H. brasiliensis rubber particles.