水稻白叶枯病菌hrpF启动子序列调控的绿色荧光蛋白基因表达体系的构建

Construction of the GFP Expression System Controlled by hrpF Promoter of Xanthomonas oryzae pv.oryzae

为了阐明启动子序列对水稻白叶枯病菌效应子基因表达的调控作用及特征,通过融合PCR将报告基因GFP基因置于水稻白叶枯病菌hrpF基因启动子序列的下游,经酶切、PCR鉴定及序列测定,成功构建了受hrpF启动子序列调控的GFP基因转录单元pMF-GFP,并转入JXO I菌株中。结果发现构建的hrpF::GFP转录单元,在hrp诱导培养基XOM2上可有效诱导表达,而在NA培养基上不能诱导表达。结果还显示,在大肠杆菌中,中间表达载体phrpF::GFP具有GFP表达活性。

In order to elucidate the expression characters for effector genes of Xanthomonas oryzae pv. oryzae （Xoo） regulated by the promoter sequence, this research put the GFP gene under the control of the promoter sequence of the Xoo hrpF gene by fusion PCR. A transcripion unit pMF-GFP was constructed and confirmed through restriction enzyme digestion, PCR and sequencing, in which the reporter gene of GFP was regulated by promoter sequence of Xoo hrpF gene. Then, the transcription uni hrpF： ：GFP was transformed to the JXO I strain. The results showed that the transcription unit could efficiently express in hrp-induced culture medium XOM2, but could not express in the NA medium. The results also showed that the GFP protein of intermediate vector phrpF ： ： GFP was obviously expressed in Escherichia coli.