Assessment of the utility of the tomato fruit-specific E8 promoter for driving vaccine antigen expression.
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.
He
Zhu-Mei
ZM
School of Life Sciences, Sun Yat-Sen University, Guangzhou, 510275, P. R. China. lsshezm@mail.sysu.edu.cn
Jiang
Xiao-Ling
XL
Qi
Yu
Y
Luo
Di-Qing
DQ
D017796:Caulimovirus / Q000235:genetics; D002772:Cholera Toxin / Q000235:genetics; D003001:Cloning, Molecular; D005638:Fruit / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D005822:Genetic Vectors / Q000138:chemical synthesis / Q000235:genetics; D006514:Hepatitis B Surface Antigens / Q000235:genetics*; D018551:Solanum lycopersicum / Q000235:genetics*; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D014026:Nicotiana / Q000235:genetics; D014170:Transformation, Genetic; D014611:Vaccination / Q000379:methods; D019444:Vaccines, DNA / Q000235:genetics*