非寄主植物烟草中与大豆疫霉菌激发素互作蛋白质的筛选

Screening of proteins interacting with Phytophthora sojae elicitins in tobacco using yeast two-hybrid system

【目的】采用酵母双杂交技术,从非寄主植物烟草中筛选与大豆疫霉菌类激发素PSE7和激发素SO-JB互作的蛋白。【方法】以PGBKT7-PSE7和PGBKT7-SOJB为诱饵质粒筛选烟草cDNA文库,得到的阳性克隆经测序后,在NCBI上进行Blast分析。【结果】PSE7和SOJB在烟草中存在几个相似的结合蛋白,表明二者具有共享的生物学功能;SOJB能与肽酶和泛素结合蛋白(与蛋白质降解有密切关系)互作,而PSE7没有筛选到类似蛋白;SOJB和PSE7均能与跨膜运输蛋白结合,PSE7与水通道蛋白结合,SOJB与铜离子转运蛋白结合,表明二者在胞质内均存在作用靶标。【结论】PSE7和SOJB最终分别从烟草中筛选出9和13个阳性互作蛋白。

[Objective] Proteins interacted with Phytophthora sojae elicitins were screened from tobacco using yeast two-hybrid system. [Method] The tobacco cDNA was screened with bait plasmid PGBKT7- PSE7 and PGBKT7-SOJB. Positive yeast clones were selected and sequenced, then analyzed with Blast in NCBI. [Result] PSE7 and SOJB had the similar binding proteins in tobacco, suggesting that they may share some similar biological functions. The ubiquitin-conjugating protein and peptidase involved in protein degradation were found binding with SOJB but not PSE7. Both SOJB and PSE7 could bind with transmem- brane transport protein,SOJB bound with copper transporter protein,and PSE7 bound with water channel protein. The results suggested that the target of SOJB and PSE7 may be located in cytoplast. [Conclusion] The results indicated that 11 proteins were interacted with PSE7 and 14 proteins interacted with SOJB.