Promoter/leader deletion analysis and plant expression vectors with the Figwort Mosaic Virus (FMV) full length transcript (FLt) promoter containing single or double enhancer domains

被引:77
作者
Maiti, IB
Ghosh, SK
Gowda, S
Kiernan, J
Shepherd, RJ
机构
[1] Department of Plant Pathology, University of Kentucky, Lexington
[2] Citrus Research and Education Center, University of Florida, 700 Experiment Station Road, Lake Alfred
[3] Dept of Tropical Public Health, Harvard School of Medicine, 665 Huntington Ave., Boston
关键词
caulimovirus; transcriptional enhancer; expression vectors; figwort mosaic virus; 35S promoter; transgenic plants; transient expression;
D O I
10.1023/A:1018477705019
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The boundaries required for maximal expression from the promoter/leader region of the full length transcript of figwort mosaic virus (FLt promoter) coupled to reporter genes were defined by 5' and 3' deletion analyses. In transient expression assays using protoplasts of Nicotiana edwardsonii, a 314 bp FLt promoter fragment sequence (-249 to +65 from the transcription start site) was sufficient for strong expression activity. Plant expression vectors developed with modified FLt promoters were tested with GUS or CAT as reporter genes in transgenic plants. The FLt promoter is a strong constitutive promoter, with strength comparable to or greater than that of the CaMV 35S promoter. The FLt promoter with its double enhancer domain linked to GUS or CAT reporter genes provides an average 4-fold greater activity than the FLt promoter with a single enhancer domain (-55 to -249 bp upstream fragment) in tests with transgenic plants and in protoplast transient expression assays.
引用
收藏
页码:143 / 156
页数:14
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