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P3N-PIPO是马铃薯Y病毒属(Potyvirus)病毒基因组中近年来新鉴定的编码蛋白。为研究P3N-PIPO在病毒与宿主互作过程中的功能,本研究以马铃薯Y病毒(potato virus Y,PVY)和马铃薯作为研究系统,用融合PCR技术扩增得到PVY编码蛋白P3N-PIPO的DNA序列,用于构建酵母双杂交诱饵质粒。为钓取马铃薯组织细胞内不同位置的互作蛋白,本研究构建了p BT3-N-P3N-PIPO、p BT3-C-P3N-PIPO、p BT3-STE-P3N-PIPO、p BT3-SUC-P3N-PIPO和p DHB1-P3N-PIPO 5个诱饵质粒,并进行自激活或毒性检测,最终获得p BT3-STE-P3N-PIPO、p BT3-SUC-P3N-PIPO和p DHB1-P3N-PIPO 3个可用于筛选马铃薯c DNA文库的诱饵质粒。
P3N-PIPO is a newly identified encoded protein in the potato genus Potyvirus. In order to study the function of P3N-PIPO in the interaction between virus and host, this study used potato virus Y (PVY) and potato as researching system, and amplified the DNA of P3Y-PIPO with PVY by fusion PCR Sequence, used to construct yeast two-hybrid bait plasmid. In order to capture the interacting proteins at different positions in potato cells, we constructed p BT3-N-P3N-PIPO, p BT3-C-P3N-PIPO, p BT3-STE-P3N-PIPO, p BT3-SUC- P3N-PIPO, and p DHB1-P3N-PIPO and were self-activated or tested for toxicity to finally obtain p BT3-STE-P3N-PIPO, p BT3-SUC-P3N-PIPO and p DHB1-P3N-PIPO3 A bait plasmid that can be used to screen potato c DNA libraries.