Cloning and DNA-binding properties of ethylene response factor, LeERF1 and LeERF2, in tomato.
Two new genes, LeERF1 andLeERF2, were isolated from a tomato (Lycopersicon esculentum cv. Lichun) cDNA library. Phylogenetic analysis indicated that they encoded Ethylene Responsive Element Binding Proteins (EREBPs), characterized by a conserved ERF (ethylene response factor) domain of specific binding plant cis-acting elements GCC box. Both LeERF1 and LeERF2 proteins were obtained via prokaryotic expression and purification. Electrophoretic mobility shift assay showed that LeERF1 and LeERF2 protein could bind to the promoter of the NP24 gene coding for pathogenesis-related protein osmotin precursor but not the mutant promoter where its GCC box was deleted. Polyclonal antibodies of LeERF1 and LeERF2 blocked their binding in vitro.
Hongxing
Zhang
Z
Department of Biotechnology, the College of Food Science and Nutritional Engineering, China Agricultural University, Beijing, 100083, China.
Benzhong
Zhu
Z
Bianyun
Yu
Y
Yanling
Hao
H
Daqi
Fu
F
Wentao
Xu
X
Yunbo
Luo
L
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D024202:Electrophoretic Mobility Shift Assay; D005030:Ethylenes / Q000378:metabolism; D018551:Solanum lycopersicum / Q000235:genetics*; D008969:Molecular Sequence Data; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism*; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*
D004268:DNA-Binding Proteins; D005030:Ethylenes; D009687:Nuclear Proteins; D010940:Plant Proteins; D014157:Transcription Factors; C036216:ethylene