Table 1.
Primers used for PCR amplification
| Primer | Sequence (5′ → 3′) a | Locations b and modifications |
|---|---|---|
| PS7-6-F |
CTAG ccatgg GA ATGAATTACACTTTAAGTG |
1aa; NcoI |
| PS7-9-R |
CG ggatcc tcacta TTAAGAATTCAGTATC |
Full-length reverse primer with stop codon; BamHI; |
| PS7-14-F |
GAACTC ggatcc ATGAATTACACTTTAAG |
1aa; BamHI; |
| PS7-15-R |
CCG ctcgag TTAAGAATTC AGTATC |
Full-length reverse primer with stop codon; XhoI; |
| PS7-16-R |
CCC aagctt ttaGTGGTGGTGGTGGTGGTG AGAATTCAGTATCTTTTTG |
Full-length reverse primer without stop codon; HindIII; six His were introduced; |
| PS7-17-R |
CCG ctcgag AGAATTCAGTATCTTTTTG |
Full-length reverse primer without stop codon; XhoI; |
| PS7-18-R |
T GC AGGAACG TCGAATGAATC |
79aa; GC substitute for AA; |
| PS7-19-F |
G CTTTTGCTGAGTTGCTTGATC |
80aa; G substitute for C; |
| TS7-23-F |
ggatcc gtcgac ccatgg GA CCCGAAATTAATCTTGTC |
25aa; BamHI, NcoI, SalI; |
| TS7-25-F |
ggatcc gtcgac ccatgg GA GTGAATTTACTATCTGAT |
44aa; BamHI, NcoI, SalI; |
| TS7-27-F |
ggatcc gtcgac ccatgg GA TTACCTTTTGCTGAGTTG |
79aa; BamHI, NcoI, SalI; |
| TS7-8-R |
CG ggatcc ggtacc tcacta TTAAGAATTCAGTATCT |
Full-length reverse primer with stop codon; BamHI, KpnI; |
| TS7-21-F |
ggatcc gtcgac ccatgg GA ATGAATTACACTTTAAGT |
1aa, BamHI, SalI, NcoI; |
| TS7-12-R |
CG ggatcc ggtacc tta ACTCTTAAATATCAAAG |
295aa; BamHI, KpnI; |
| TS7-14-R |
CG ggatcc ggtacc tta AAACAGAGAATACCAATAATC |
287aa; BamHI, KpnI; |
| TS7-16-R |
CG ggatcc ggtacc tta GGGTATCATACTTAATTTTCC |
214aa; BamHI, KpnI; |
| MSKP1-1-F |
CG ggatcc ATGGCCGCCGAGGGC |
1aa; BamHI; |
| MSKP1-2-R |
CCG ctcgag CTCGAAGGCCCACTG |
Full-length reverse primer without stop codon; XhoI; |
| MSKP1-3-F |
CG gaattc ATGGCCGCCGAGGGC |
1aa; EcoRI; |
| MSKP1-4-R |
CG ggatcc TACGTGCTTGTTGCAG |
67aa; BamHI; |
| MSKP1-5-F |
CG gaatcc GAGGACCTCAAGAAC |
97aa; EcoRI; |
| MSKP1-6-R |
CG ggatcc ACCCTTGATGTTCAGATAG |
128aa; BamHI; |
| MSKP1-8-F |
CCG ctcgag CT ATGGCCGCCGAGGG |
1aa; XhoI; |
| MSKP1-9-R |
CG ggatcc CTCGAAGGCCCACTG |
Full-length reverse primer without stop codon; BamHI; |
| MSKP1-10-F | CCC aagctt CG ATGGCGGCCGAGGGCGAG | 1aa; HindIII; |
aIntroduced restriction endonuclease sites are in lower case. Two extra nucleotides (italicized) were added to allow in-frame expression of fusion proteins of interest. The italicized nucleotides in lower case denote the stop codon added, while three mutant sites are denoted by boldfaced capitals in italic.
bNumbered according to the amino acid sequence of P7-2 or SKP1Maize. The F or R designation in the primer names denotes that the primer is a forward (5′) or reverse (3′) primer, respectively.