Supplementary Materials
Characterization of the Scutellaria barbata glycosyltransferase gene and its promoter
Planta
Shu-Jiau Chiou · Wen-Yi Liu · Cheng-Li Fang · Tsai-Yun Lin
Tsai-Yun Lin ()
Department of Life Science and Institute of Bioinformatics and Structural Biology, National Tsing Hua University, No. 101, Sec. 2, Kuang Fu Rd., Hsinchu, 30013 Taiwan, R.O.C.
E-mail: tylin@life.nthu.edu.tw
Phone: +886-3-574-2578
Fax: +886-3-571-5934
Supplementary Table 1 PCR primers used in this study
(A) Primers designed for genome walking
|
Sequence1
|
Position2
|
Construct
(pCR2.1 or pBI121)
|
SbP-GSP1
|
5'- TCATTGGAACAAGGATAATATGTAGCT -3'
|
+119 to +145
|
pCR-SbP
SbP-850U
|
SbP-GSP2
|
5'- TGTAAAATGGAAAGATCTTAATTATG -3'
|
+78 to +103
|
(B) Primers designed for serial deletions
|
|
|
|
SbP-F1
|
5'- GTGTCCAATTCCTGCAGGCTCTAATCTAG -3'
|
127 to 98
|
pCR-109
SbP-109
|
SbP-F3
|
5'- TAAGAGTATCCTGCAGGAATGTAGGAAT -3'
|
216 to 198
|
pCR-215
SbP-215
|
SbP-F4
|
5'- CATAAACTAACCTGCAGGCAATCCAAC - 3'
|
262 to 235
|
pCR-250
SbP-250
|
SbP-F5
|
5'- TACATTAATCCTGCAGGACACACCCTATTTT -3'
|
403 to 381
|
pCR-392
SbP-392
|
SbP-BamR
|
5'- TGGCCTGCATGAGAAAGGATCCAATATAGGAT -3'
|
+33 to +4
|
|
SbP-D5-F
SbP-D5-R
|
5'- CCAACTTTTTTTTCAATTTGACTC -3'
5'- GAGTCAAATTGAAAAAAAAGTTGG -3'
|
240 to 228
177 to 167
|
pCR-39262
SbP-39262
|
(C) Primers designed for intron detection
|
|
|
|
SbUGT-full-F
SbUGT-in-R
SbUGT-in-F
SbUGT-full-R
|
5'-TGAAATGGGACAGCTACATATTATC-3'
5'-TTCCTTCCAAAGTCGAGTTCCATCCACA-3'
5'-TCAACTCAAAAAAGCCCAACTCAGTTGT-3'
5'-CTAGTTTATGGCCAGTTTCATA-3'
|
+107 to +132
+1186 to +1213
+920 to +947
+1524 to +1545
|
|
(D) Primers designed for RT-PCR and real time RT-PCR
|
|
|
|
SbGT-RT-F
SbGT-RT-R
SbGT-qRT-F
SbGT-qRT-R
Actin-F
Actin-R
|
5'-ATTGCCAGACAACATATTGAG-3'
5'-GTGGAAGATTCGGAATGAC-3'
5'-TTCTTGCCATGGACGACTG-3'
5'-ATGGAAGATCAACCTCGGAAT-3'
5'-TCACCATTGGAGCTGAGAGATTC-3'
5'-CTGCAGCTTCCATTCCAATCA-3'
|
+207 to +227
+493 to +511
+458 to +475
+496 to +516
|
|
1Nucleotides changed to create a restriction endonuclease cleavage site are underlined
2Position is given relative to the putative SbUGT transcription start site
Supplementary Table 2 Cis-acting regulatory elements in SbUGT promoter.
The SbUGT promoter sequence was screened against PlantCARE and PLACE databases. The positions are given relative to the putative transcription start site of the SbUGT promoter
Name
|
Position
|
Sequence
|
Putative function
|
Reference
|
Pyrimidine box
|
-148 (+)
-53 (+)
-440 (+)
-727 (-)
|
TTTCTTCTCT
TTTTTTCC
|
Cis-acting element conferring high transcription levels
GA and ABA-responsive element
|
Daraselia et al. (1996)
Cercós et al. (1999)
|
ABRE
|
-122 (-)
-956 (+)
|
GGACACGTGGC
CACGTG
|
Cis-acting element involved in the abscisic acid responsiveness
|
Hobo et al. (1999)
|
MYB motif
|
-771 (+)
-548 (+)
-546 (-)
|
CNGTTR
|
Regulate genes responsive for plant development, flavonoid biosynthesis and ABA
|
Romero et al. (1998)
Wu et al.. (2006)
|
CE3
|
-132 (+)
|
GACGCGTGTC
|
Cis-acting element involved in ABA and VP1 responsiveness
|
Hobo et al. (1999)
|
AT1-motif
|
-613 (-)
|
ATTAATTTTACA
|
Light responsive module
|
Datta and Cashmore (1989)
|
Box-W1
|
-918 (-)
|
TTGACC
|
Fungal elicitor responsive element
|
Rushton et al. (1998)
|
G-Box
|
-131 (+)
-956 (±)
-946 (+)
|
ACACGTGT
CACGTG
CACGAC
|
Cis-acting regulatory element involved in light responsiveness
|
Safrany et al. (2008)
|
I box
|
-91 (+)
-188 (+)
-318 (+)
-720 (-)
|
GATAA
|
Light-responsive
|
Fiebig and Link (1992)
|
CAAT-box
|
+63 (-)
+60 (+)
-62 (+)
-171 (-)
-286 (+)
-365 (+)
-416 (+)
-433 (+)
-579 (-)
-720 (+)
|
CAAAT
|
Cis-acting element in promoter and enhancer regions
|
Rieping and Schöffl (1992)
|
CGTCA-motif
|
-368 (+)
-68 (+)
|
CGTCA
|
Cis-acting regulatory element involved in the MeJA-responsiveness
|
Rouster et al. (1997)
|
ELI-box3
|
-154 (-)
|
AAACCAATT
|
Elicitor-responsive element
|
|
GARE motif
|
-35 (-)
|
AAACAGA
|
Gibberellin-responsive element
|
Ogawa et al. (2003)
|
GCN4 motif
|
-176 (-)
|
TGAGTCA
|
Cis-regulatory element involved in endosperm expression
|
Washida et al. (1999)
|
MBS motif
|
-766 (-)
|
CAACTG
|
MYB binding site involved in drought
|
Simpson et al. (2003)
|
TCA element
|
-240 (+)
|
CCATCTTTTT
|
Cis-acting element involved in salicylic acid responsiveness
|
Rushton et al. (1998)
|
W box
|
-172 (+)
-182 (+)
-701 (+)
-814 (-)
|
TGACT
|
Sugar and/or wound-inducible element
|
Sun et al. (2003)
|
WB box
|
-174 (+)
-818 (-)
|
TTTGACY
|
Responsive to abiotic and biotic stresses, biosynthetic pathway and hormone signaling
|
Eulgem et al. (2000)
|
Amylase box
|
-476 (+)
|
TATCCAT
|
Sugar response element
|
Lu et al.. (1998)
|
A box
|
-82 (+)
|
TACGTA
|
Sugar-repression element
|
Toyofuku et al. (1998)
|
TATCCAT/C motif
|
-750 (+)
|
GAGAAGAATA
|
Sugar-responsive
|
Lu et al.. (1998)
|
GT1 box
|
-312 (+)
-434 (-)
-605 (-)
-733 (+)
|
GAAAAA
|
NaCl and pathogen-inducible element
|
Park et al. (2004)
|
AGAAA motif
|
+18 (-)
-144 (-)
-313 (+)
-462 (+)
-619 (-)
-660 (+)
-670 (-)
-851 (-)
-858 (-)
-902 (-)
|
AGAAA
|
Pollen-specific
|
Bate and Twell (1998)
|
Root motif
|
-26 (+)
-293 (±)
-362 ±)
-377 (-)
-427(-)
-585 (+)
+7 (+)
+40 (±)
+74 (-)
|
ATATT
|
Root-specific
|
Elmayan and Tepfer (1995)
|
OSE2
|
-169 (+)
-221 (-)
-461 (-)
-516 (-)
|
CTCTT
|
Root-specific expression
|
Stougaard et al. (1990)
|
RY repeat motif
|
+23 (+)
|
CATGCA
|
Seed-specific
|
Bäumlein et al. (1992)
|
NRR motif
|
-1072 (-)
|
TAGTGGAT
|
Stress negative regulatory element
|
Elliott and Shirsat (1998)
|
E box
|
-130 (+)
-155 (+)
-763 (+)
-771 (+)
-956 (+)
|
CANNTG
|
MYC-recognition site
Tissue-specific
Light-responsive
|
Hartmann et al. (2005)
|
KCACGW
|
-124 (-)
-947 (±)
-1023 (+)
|
|
Root hair-specific cis-elements
|
Shangguan et al. (2008)
|
References
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