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通过Tail PCR分离了GHNBS基因的5’侧翼序列,PLACE分析表明其含有CAAT-box、TATA-box及乙烯、茉莉酸甲酯、脱落酸应答元件,病原菌诱发子反应元件W boxes、GT-1、MYB、MYBST1和MYB1LEPR等。同时,在这段序列上还存在一些根特异表达元件。包括2个as-1和1个EECCRCAH1在内的3个增强子元件也存在于这段序列的上游区域,它们可能增强GUS基因在转基因植物中的表达。将GHNBS基因的5′调控序列以不同缺失与GUS基因融合转化拟南芥,发现GUS基因主要在根、茎的韧皮部和叶脉中表达。PGN-1559和PGN-1117表现为器官特异性表达。而PGN-476不能特异表达,同时也不能在根部表达。SA,ABA,MeJA,Eth-ylene,枯萎病菌和细菌DC3000处理后GUS活性均有显著上升,说明GHNBS基因的5′调控序列含有相应的应答反应因子,是一个器官特异以及与病原相关的启动子。
The 5 ’flanking sequence of GHNBS gene was isolated by Tail PCR and the PLACE analysis indicated that it contained CAAT-box, TATA-box and ethylene, methyl jasmonate, abscisic acid response element, pathogen elicitor reaction element W boxes, GT- MYB, MYBST1 and MYB1LEPR and so on. At the same time, there are some root specific expression elements in this sequence. Three enhancer elements, including two as-1 and one EECCRCAH1, are also present in the upstream region of this sequence, which may enhance the expression of the GUS gene in transgenic plants. Arabidopsis thaliana was transformed with the GUS gene by 5 ’regulatory sequence of GHNBS gene. The GUS gene was mainly expressed in the phloem and veins of roots and stems. PGN-1559 and PGN-1117 showed organ-specific expression. PGN-476 can not be expressed specifically, nor can it be expressed at the roots. The GUS activity of SA, ABA, MeJA, Eth-ylene, Fusarium oxysporum and bacterial DC3000 increased significantly, indicating that the 5 ’regulatory sequence of GHNBS gene contains the corresponding response factor and is an organ-specific and pathogen-related promoter .