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从拟南芥JAtYTAC基因组文库中完整地亚克隆了FLS2和LysM两个抗病基因,亚克隆的DNA片段中分别包含各自的启动子、内含子和终止子。利用农杆菌介导的方法分别将这两个基因转化番茄品种‘Moneymaker’,转基因植株经PCR初步筛选后再分别提取总RNA进行RT-PCR,以验证其是否在‘Moneymaker’中表达。试验结果表明,FLS2和LysM均能成功地在‘Moneymaker’中表达,说明番茄的转录系统不仅能识别这两个抗病基因的启动子并起始转录,还能正确识别并剪切这两个基因的外显子和内含子。据此推测,拟南芥和番茄这两个远缘物种的转录机制存在相对保守的成份,这为探索在这两个物种间R基因的功能及其下游信号通路的保守性提供了有价值的参考。
FLS2 and LysM genes were completely subcloned from the Arabidopsis JAtYTAC genomic library. The subcloned DNA fragments contained their respective promoters, introns and terminators. The two genes were transformed into ’Moneymaker’ by Agrobacterium-mediated transformation. The transgenic plants were initially screened by PCR and then total RNA was separately extracted for RT-PCR to verify whether they were expressed in ’Moneymaker’. The results show that both FLS2 and LysM can be successfully expressed in ’Moneymaker’, indicating that the tomato transcription system can not only recognize the promoter of the two disease-resistance genes and initiate transcription, but also correctly recognize and cut the two Gene exons and introns. Therefore, it is speculated that the relative conservatism of the transcriptional mechanisms of Arabidopsis and tomato, two distant species, provides valuable information for exploring the function of the R gene and the conservatism of downstream signaling pathways between these two species reference.