摘要
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 was a newly identified protein encoded by viral genome of potyvirus in recent years. To study the function of P3N-PIPO during the virus-host interaction process, potato virus Y （PVY） and potato were used as the research system. The DNA sequence of PVY P3N-PIPO was amplified by fusion PCR technology, which was used to construct the bait plasmids in yeast two-hybrid system. In order to fish interacting proteins from different subcellular fractions of potato celluar tissues, pBT3-N-P3N-PIPO, pBT3-C-P3N-PIPO, pBT3-STE-P3N-PIPO, pBT3-SUC-P3N-PIPO and pDHB I-P3N-PIPO five altemative bait plasmids were constructed, and self-activations and toxicities of them were tested subsequently. Ultimately, pBT3-STE-P3N-PIPO, pBT3-SUC-P3N-PIPO and pDHB 1-P3N-PIPO three bait plasmids were proved to be available for screening the interacted proteins from the cDNA library of PVY-infected potato.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2017年第1期299-304,共6页
Genomics and Applied Biology
基金
国家自然科学基金(31260030)
广西回国基金重点项目(2013GXNSFCB019003)共同资助
