| 摘要: |
| [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SOJB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. |
| 关键词: 大豆疫霉菌 激发素 酵母双杂交系统 烟草 蛋白质筛选 |
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| 基金项目:国家重点基础研究发展规划(973计划)，教育部长江学者和创新团队发展计划，高等学校学科创新引智计划项目 |
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| Screening of proteins interacting with Phytophthora sojae elicitins in tobacco using yeast two-hybrid system |
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| Abstract: |
| 【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 transmembrane 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. |
| Key words: Phytophthora sojaeelicitin yeast two hybrid system tobacco protein screen |
