Supplementary Data
Isolation and characterization of a gene encoding an ethylene responsive factor (ERF) protein from Ceratoides arborescens
Jie Dong1†, Xuemin Wang1†, Kang Wang2,Zan Wang1，Hongwen Gao1*
1 Institute of Animal Science, ChineseAcademy of Agricultural Sciences,
2-Yuan-Ming-Yuan West Rd., Haidian District, Beijing, 100193, People’s Republic of China
2 Institute of Grassland Science, China Agricultural University,
2-Yuan-Ming-Yuan West Rd., Haidian District, Beijing, 100193,People’s Republic of China,
SupplementaryTable 1
Table 1 The primes used in gene clone
Primers / The Sequences of Primers (5’-3’)AP2-up / 5’-GGNAAGTGGGTBGSTGAAAT-3’
AP2-low / 5’-CATCATANGCDAGNGCAGC-3’
GSP1 / 5’-GCACTCTCGGGAGGGGTAGTAAAG-3’
GSP2 / 5’-CGCTGAAGAAGCTGCTAGAGCATAC-3’
UPM / 5’-CTAATACGACTCACTATAGGGCAAGCAAGCAGTGGTATCAACGCAGAGT-3’
CeERFover-up / 5’-ATGTGTGGAGGAGCAGTAATCTC-3’
CeERFover-low / 5’-TCAGAAGACTCCCCCAGGGAAGTCGT-3’
SpeI-up / 5’-CGGACTAGTATGTGTGGAGGAGCAGTAATCTC-3’
SpeI-low / 5’-CGGACTAGTGAAGACTCCCCCAGGGAAGTCGT-3’
SupplementaryFig. 1
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Figure Legends
SupplementaryFig.1The full nucleotide and deduced amino acid sequences of CeERF gene.
Note: The AP2/ERF domain was underlined and the acidic domain was indicated with the underline. The codonsof start and stop were indicated by shadow. Black box represents MCGGAVI motif. The alanine and aspartic acid residues were marked by asterisks. The acidic amino acid region was showed by double underline, and the nuclear localization signal was showed by dotted line.
SupplementaryFig.2 Phylogenetic analysis of CeERF with other AP2/ERF transcription factor proteins in 59 other plant species. Phylogenetic tree shows the relationship between CeERF and other ERFs proteins.
SupplementaryFig. 3Multiple alignment analysis of CeERF with other ERFs from five legume plants. Multiple sequence alignments of the predicted amino acid sequence of CeERFwith proteins in GenBank with the highest homology. The amino acid sequences of Ceratoides arborescensCeERF (ACI62768), Galegae orientalisGoDREB (FJ223566),Medicago sativa MsERF (ABO93372),Trifolium pratenseTpERF (BAE71206),Glycine max GmERF (AAQ10777) and Arabidopsis thalianaAtERF (AAM63284)were aligned.
SupplementaryFig. 4 Root growth of transgenicand control tobacco seedling under different hormone conditions. Transgenic and control lines were treated with ABA,JA and SA, respectively.Each treatment was set four concentrations (0, 100, 200 and 500µM). Means (±S.E.)of three replicates.