Characterization and activity enhancement of the phloem-specific pumpkin PP2 gene promoter.
The promoter of the pumpkin (Cucurbita moschata) PP2 gene (designated NP) was isolated from the restriction enzyme-digested genomic DNA pool by genome walking and its activity and phloem specificity were examined in transgenic tobacco plants by using GUS as a reporter. Deletion analysis of the promoter revealed that the 473-bp fragment (-465 to + 8 relative to the transcription start site; designated as NPII) exhibited similar activity as the full-length NP promoter and retained its phloem specificity. Furthermore, the sequence from -465 to -171 was shown to contain positive regulatory cis-elements for the promoter activity. An enhanced NP promoter was constructed by duplicating the sequence -465 to -85, and its activity in phloem tissue was shown to be higher than that of the Commelina Yellow Mottle Virus (CoYMV) promoter or a chimeric promoter consisting of the double enhancer sequence from the Cauliflower Mosaic Virus (CaMV) 35S promoter fused upstream to the NPII fragment.
Guo
Hongnian
H
National Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.
Chen
Xiaoying
X
Zhang
Haili
H
Fang
Rongxiang
R
Yuan
Zhengqiang
Z
Zhang
Zhenshan
Z
Tian
Yingchuan
Y
D016960:Agrobacterium tumefaciens / Q000235:genetics; D017796:Caulimovirus / Q000235:genetics; D003001:Cloning, Molecular; D034822:Commelina / Q000821:virology; D028464:Cucurbita / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D017353:Gene Deletion; D017343:Genes, Plant; D017930:Genes, Reporter; D005822:Genetic Vectors; D008156:Luciferases / Q000378:metabolism; D037121:Plant Lectins / Q000235:genetics*; D010942:Plant Viruses / Q000235:genetics; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D014026:Nicotiana / Q000235:genetics*; D014170:Transformation, Genetic
D037121:Plant Lectins; C091028:phloem lectin; D008156:Luciferases