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ONLINE RESOURCE
Molecular and biochemical identification of inositol 1,3,4,5,6-pentakisphosphate 2-kinase encoding mRNA variants in castor bean (Ricinus communis
L.) seeds, Planta, Jaeju Yu, Adolfo Saiardi, John S. Greenwood* and J. Derek Bewley
* Corresponding author:
Department of Molecular and Cellular Biology
University of Guelph, Guelph, Ontario, N1G 2W1 Canada
E-mail: jgreenwo@uoguelph.ca
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Online Resource 1. A list of synthesized primer sequences described in the text of this study. Underlined regions indicate restriction enzyme sites or histidine
encoding sequences.
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Name
Oligonucleotide sequence (5’-3’) and Description
50F
CTAGATCATATGGAGGTCAAATTAGAGAAAAAGG
51R
TAATTTCTCGAGCAAGCAAACGTCGCATCTC
Amplification and cloning RcIPK1 cDNA with restriction enzyme sites, NdeI and XhoI
57F
TAATACGACTCACTATAGGGG
58R
CTAGTTATTGCTCAGCGG
Sequencing inserts cloned in pET-22b(+) vector
Sac67F
ACGTGAGCTCATGGAGGTCAAATTAGAGAAAAAGG
Not73R555
ACGTGCGGCCGCCTACAAGCAAACGTCGCATCTCTG
Amplification and cloning DS46 RcIPK1 variant with restriction enzyme sites, SacI and NotI
Sac67F
ACGTGAGCTCATGGAGGTCAAATTAGAGAAAAAGG
Not72R452
ACGTGCGGCCGCCTATATAGTTTCACAAACCTCCATATTCTC
Amplification and cloning DB39 RcIPK1 variant with restriction enzyme sites, SacI and NotI
Sac67F
ACGTGAGCTCATGGAGGTCAAATTAGAGAAAAAGG
Not71R450
ACGTGCGGCCGCTTATTTATTGCTCTCCATATTCTCGTTG
Amplification and cloning AB56 RcIPK1 variant with restriction enzyme sites, SacI and NotI
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Sac67F
ACGTGAGCTCATGGAGGTCAAATTAGAGAAAAAGG
Not70R277
ACGTGCGGCCGCTCATCTGCCTGGATAAATCCC
Amplification and cloning DB37 RcIPK1 variant with restriction enzyme sites, SacI and NotI
Sac67F
ACGTGAGCTCATGGAGGTCAAATTAGAGAAAAAGG
Not69R224
ACGTGCGGCCGCTCAATGCATCTTCAAAACCTTGTC
Amplification and cloning AS14 RcIPK1 variant with restriction enzyme sites, SacI and NotI
Sac67F
ACGTGAGCTCATGGAGGTCAAATTAGAGAAAAAGG
Not68R215
ACGTGCGGCCGCTCATATTGGCTTACCTCCGATG
Amplification and cloning AB55 RcIPK1 variant with restriction enzyme sites, SacI and NotI
Sac67F
ACGTGAGCTCATGGAGGTCAAATTAGAGAAAAAGG
Not73R555His
ACGTGCGGCCGCCTAGTGGTGGTGGTGGTGGTGCAAGCAAACGTCGCATCTCTG
Amplification and cloning DS46 RcIPK1 variant with restriction enzyme sites, SacI and NotI and 6 histidine encoding sequences
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ONLINE RESOURCE
Molecular and biochemical identification of inositol 1,3,4,5,6-pentakisphosphate 2-kinase encoding mRNA variants in castor bean (Ricinus communis
L.) seeds, Planta, Jaeju Yu, Adolfo Saiardi, John S. Greenwood* and J. Derek Bewley
* Corresponding author:
Department of Molecular and Cellular Biology
University of Guelph, Guelph, Ontario, N1G 2W1 Canada
E-mail: jgreenwo@uoguelph.ca
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Online Resource 2. Multiple sequence alignment of RcIPK1 genes. Following sequencing analysis, the corresponding cDNA sequences from AB (3, 10, 55, and
56), CB (24, 25, 27, and 31), DB (33, 37, 39, and 40), AS (14, 17, 22, and 80), DS (46, 50, 52, and 106) were aligned and grouped. Six mRNA sequences
representative of each group, RcIPK1A: the RcIPK1 genomic DNA sequence (NCBI accession number: NW_002994277.1), and RcIPK1B: the RcIPK1 mRNA
sequence (NCBI accesssion number: XM_002510379.1) were aligned using DNAMAN 4.1.1.1 software (Lynnon Biosoft). Eleven exons are indicated by
nucleotides in bold. Retained introns, deleted exons and nucleotides inserted into exons are represented by underlining, dashed spaces, and circles, respectively.
Different sequences are in lower case letters. Each termination codon is indicated by boxes.
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