He Zhu-Mei, Jiang Xiao-Ling, Qi Yu, Luo Di-Qing
School of Life Sciences, Sun Yat-Sen University, Guangzhou, 510275, P. R. China.
Genetica. 2008 Jun;133(2):207-14. doi: 10.1007/s10709-007-9201-2. Epub 2007 Sep 5.
To assess the utility of the tomato fruit-specific E8 gene's promoter for driving vaccine antigen expression in plant, the 2.2 kb and 1.1 kb E8 promoters were isolated and sequenced from Lycopersicon esculentum cv. Jinfeng #1. The 1.1 kb promoter was fused to vaccine antigen HBsAg M gene for the transfer to Nicotiana tabacum, and the CaMV 35S promoter was used for comparison. Cholera toxin B (ctb) gene under the control of the 1.1 kb promoter was transformed into both N. tabacum and L. esculentum. Southern blot hybridization confirmed the stable integration of the target genes into the tomato and tobacco genomes. ELISA assay showed that the expression product of HBsAg M gene under the control of the 1.1 kb E8 promoter could not be detected in transgenic tobacco tissues such as leaves, flowers, and seeds. In contrast, the expression of HBsAg M gene driven by CaMV 35S promoter could be detected in transgenic tobacco. ELISA assay for CTB proved that the 1.1 kb E8 promoter was able to direct the expression of exotic gene in ripe fruits of transgenic tomato, but expression was absent in leaf, flower, and unripe fruit of tomato, and CTB protein was not detected in transgenic tobacco tissues such as leaves, flowers, and seeds when the gene was under the control of the 1.1 kb E8 promoter. The results indicated that the E8 promoter acted not only in an organ-specific, but also in a species-specific fashion in plant transformation.
为评估番茄果实特异性E8基因启动子在驱动植物中疫苗抗原表达方面的效用,从番茄品种金丰1号中分离并测序了2.2 kb和1.1 kb的E8启动子。将1.1 kb启动子与疫苗抗原HBsAg M基因融合,用于转化烟草,同时以花椰菜花叶病毒35S启动子(CaMV 35S启动子)作为对照。将受1.1 kb启动子控制的霍乱毒素B(ctb)基因转化到烟草和番茄中。Southern杂交证实目标基因稳定整合到番茄和烟草基因组中。ELISA检测表明,在转基因烟草的叶片、花和种子等组织中,未检测到受1.1 kb E8启动子控制的HBsAg M基因的表达产物。相比之下,在转基因烟草中可检测到受CaMV 35S启动子驱动的HBsAg M基因的表达。CTB的ELISA检测证明,1.1 kb E8启动子能够指导外源基因在转基因番茄的成熟果实中表达,但在番茄的叶片、花和未成熟果实中不表达,并且当该基因受1.1 kb E8启动子控制时,在转基因烟草的叶片、花和种子等组织中未检测到CTB蛋白。结果表明,E8启动子在植物转化中不仅具有器官特异性,还具有物种特异性。