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本文通过片段缺失和定点突变的方法构建产生水稻(Oryza sativa)水通道蛋白OsPIP1;3的变异本。通过膜酵母双杂交进行蛋白互作分析,实验结果表明水稻白叶枯病菌(Xanthomonas oryzae pv.oryzae)harpin蛋白Hpa1能与OsPIP1;3互作,但如果将OsPIP1;3的一个胞外序列(loopE)及OsPIP1;3的第六个跨膜区域(TM6)删除,则互作不能发生。进一步对276~279 aa进行替换,OsPIP1;3与Hpa1不再发生互作。这些结果说明,OsPIP1;3序列上的loop E和TM6上的氨基酸对OsPIP1;3–Hpa1互作有重要影响,而276~279 aa是可能的互作位点。
In this study, a variant of OsPIP1; 3, which produces aquaporin (Oryza sativa), was constructed by fragment deletion and site-directed mutagenesis. The interaction between OsPIP1; 3 and Hpa1 in Xanthomonas oryzae pv. Oryzae was elucidated by membrane-yeast two-hybrid assay. However, if an extracellular sequence of OsPIP1; 3 ) And the sixth transmembrane region (TM6) of OsPIP1; 3, the interaction can not occur. 276 ~ 279 aa further replacement, OsPIP1; 3 and Hpa1 no longer occur interaction. These results indicate that the amino acids at loop E and TM6 on OsPIP1; 3 sequence have an important effect on OsPIP1; 3-Hpa1 interaction, while 276 ~ 279 aa are possible interaction sites.