Supplemental table 1. Primers used in the paper
Constructions
Restriction sites
Primer sequences
Primers of amplifing ZmAN13 for southern blot and semi-quantitative PCR
ZmAN13 fragment sth-F
5' ATGGAGCACAAGGAGGCTGG 3'
ZmAN13 fragment sth-R
5' CTAGATCTTGTCGAGCTTGTCTGC 3'
Primers of amplifing ZmAN13 fragment for Real-time PCR
ZmAN13 fragment realtime-F
5' CGGTAGTAGCTGTTGCCCAA 3'
ZmAN13 fragment realtime-R
5' TACAAGTTCCCACACCGGC 3'
Primers of amplifying α-tubulin fragment for Real-time PCR
Tubulin-F
5' GCTATCCTGTGATCTGCCCTGA 3'
Tubulin-R
5' CGCCAAACTTAATAACCCAGTA 3'
Primers of amplifying actin fragment for semi-quantitative PCR
Actin-F
5' CATCAGGAAGGACTTGTACGG 3'
Actin-R
5' GATGGACCTGACTCGTCATAC 3'
Primers of creating ZmAN13 subcellure localization vector
pMDC85-ZmAN13-F
SpeI
5' CACTAGTATGGAGCACAAGGAGGCT 3'
pMDC85-ZmAN13-R
KpnI
5' CGGTACCGAGATCTTGTCGAGCTTGT 3'
pUCLNGFP2-ZmAN13-F
KpnI
5' GGGTACCCATGGAGCACAAGGAGG 3'
pUCLNGFP2-ZmAN13-R
SacI
5' GGAGCTCCTAGATCTTGTCGAGCT 3'
Primers of creating pDEST32 vector
pDEST32-ZmAN13-F
5' ATGGAGCACAAGGAGGCTGG 3'
pDEST32-ZmAN13-R
5' CTAGATCTTGTCGAGCTTGTCTGC 3'
pDEST32-ZmERF11-F
5' CCAACCTTTCCGCCACTC 3'
pDEST32-ZmERF11-R
5' CAGGATCATCGCATAGACCAG 3'
Primes of creating ZmAN13 overexpression vector
p3301-ZmAN13-F
XbaI
5' GTCTAGAATGGAGCACAAGGAGGCT 3'
p3301-ZmAN13-R
BamHI
5' GGGATCCCTAGATCTTGTCGAGCTT 3'
Primers of creating yeast Two-Hybrid Assays vector
pGBKT 7-ZmAN13-F
NcoI
5' TCCATGGAGCACAAGGAGGCTGGGT 3'
pGBKT 7-ZmAN13-R
BamHI
5' TGGATCCCTAGATCTTGTCGAGCTT 3'
pGADT 7/ pGBKT 7-A20-F
NdeI
5' CCATATGGCCCCCGAGGGACCCA 3'
pGADT 7/ pGBKT 7-A20- -R
BamHI
5' CGGATCCATCCTGCTTCGTTATCA 3'
pGADT 7/ pGBKT 7-AN1-F
NdeI
5' CCATATGTCCAAAGGGGGGAAGGT 3'
pGADT 7/ pGBKT 7-AN1-R
BamHI
5' CGGATCCCCGGTAGTCGAACTTGC 3'
Supplemental Figure 1
Fig. S1. Sequence analyses of ZmAN13.
(A) Phylogenetic relationship of ZmAN13 with other A20/AN1 genes from maize or
rice. Multiple sequence alignment was performed using the program ClustalX1.83,
and the phylogenetic tree was constructed using the program treev32. The proteins
accession numbers in the GeneBank or Swiss-prot are: ZmAN11, DQ244747;
ZmAN14, EF396226; OsSAP4, AK067403; OsSAP8, AAQ84334.1.
(B) Conservation of the amino acids at N-terminal A20 type zinc-finger domain and
the C-terminal AN1 type zinc-finger domain of ZmAN13 with OsSAP8
(AAQ84334.1), ZmAN11 (DQ244747), ZmAN14 (EF396226), ZFAN5 (ZnF216) of
humans (AAC61801.1) and mouse (AAC42600.1), ZFAN6 of human (AAH05283)
and rat (AAH76394), Conserved positions of cysteine and histidine residues were
marked by asterisk (*). Multiple sequence alignment was performed using the
program ClustalX1.83.
Supplemental Figure 2
Fig. S2. Trans-activation potential analysis of ZmAN13 protein in yeast cells.
The transformants were spoted on the SD/-Leu and SD/-Leu-His medium respectively,
for examination of growth.