1
Plant Molecular Biology Reporter
SUPPLEMENTARY TABLE
Identification and molecular characterization of a lectin
receptor-like kinase (GhLecRK-2) from cotton
Sonia M Phillips1, Ian A Dubery1*, Henriette van Heerden 1, 2
1Department
of Biochemistry, University of Johannesburg, Kingsway Campus, P.O. Box 524, 2006, Auckland Park,
Johannesburg, South Africa
2Current
address: Department of Veterinary Tropical Diseases, University of Pretoria, Private Bag X04, Onderstepoort,
0110, South Africa
2
Plant Molecular Biology Reporter
Table S1-S5: Gene-specific primers used in the genome walking reactions, 5’ and 3’ RACE reactions,
internal PCRs, Southern blots and RT-qPCR for the molecular characterization of GhLecRK2.
Name
S1
Sequence (5’ – 3’)
Details
Tm
GC %
Length
(nt)
primary PCR
secondary PCR
65
65
60
57
20
21
primary PCR
secondary PCR
60
58
50
45
20
20
Genome walking primers
Upstream Genome walk
R1
R2
CCGCTGCTCATGGAACTGGA
CGTTCATCCCCGTTGAGAAGC
Downstream Genome Walk
F1
F2
S2
TCGCTGGTTTCTCTTCCAGT
CTCATTGATTGGGTTGCTGA
5’ and 3’ RACE primers
First 5’ RACE
RR3
TCAGCAACCCAATCAATGAG
cDNA synthesis
58
45
20
RR2
CCGCTGCTCATGGAACTGGA
primary PCR
65
60
20
RR1
ACTGGAAGAGAAACCAGCGA
secondary PCR
60
50
20
Second 5’ RACE
5’Exon2R3
TTGCCATTGCGAGCCAGTT
cDNA synthesis
60
53
19
LecDR1
AAAGAAGAAGATTTGGGAGAAGGTG
primary PCR
61
40
25
LecDR2
AGGTGAAGACTTGTCGTCCAA
secondary PCR
60
48
21
NF1
GATAACTGGGAGGCGTTTGA
primary PCR
60
50
20
F1
TCGCTGGTTTCTCTTCCAGT
secondary PCR
60
50
20
58
58
45
45
20
20
58
58
45
45
20
20
3’ RACE
S3 Internal PCR primers
PCR vs. RT-PCR of C4B5 EST
QPKF1
RR3
AAAGCCAGTTGATTCGAGGA
TCAGCAACCCAATCAATGAG
RT-PCR for further cDNA sequence
LecD2F
RR3
GCCCACAAATTTCATCTTCC
TCAGCAACCCAATCAATGAG
S4 Southern blot primers
LecD 2F
GCCCACAAATTTCATCTTCC
58
45
20
LecD R1
AAAGAAGAAGATTTGGGAGAG
61
40
25
S5 RT-qPCR primers
LF1
ATATTGGAGAGCGGTGATGC
60
50
20
LR1
GCTCGACTTTTGTCCCGTAG
62
55
20