TABLE 2.
Primers used for PCR, mutagenesis, and 5′ RACE PCR experimentsa
| Name | Sequence | Characteristic |
|---|---|---|
| S1 | 5′-CAAATTCtGCaGCGTTTAGAAAC-3′ | (+) PstI |
| S2 | 5′-CTTCGGAGcTCATTCAAACTG-3′ | (−) SacI |
| S3 | 5′-ggatcccgggAAAtGATGAACATGTTTT AGCC-3′ | (+) SmaI |
| S4 | 5′-ggatcccgggTTCaAATATGAACAGCTT CTCG-3′ | (−) SmaI |
| S7 | 5′-CGTCCTGTTGATTGGTCGC-3′ | (+) |
| S8 | 5′-CCATTGCCATTCGCCCTTTC-3′ | (−) |
| S12 | 5′-GTCGATTGGtaCCTGAGATTAC-3′ | (+) KpnI |
| S1B | 5′-GGAAAGGAGGaTCCAGAAATC-3′ | (+) BamHI |
| S2B | 5′-CCATTATTTTTTCGgaTCCTTTC-3′ | (−) BamHI |
| AS1 | 5′-CAACGGATCTAgAtAGCGGAAG-3′ | (+) XbaI |
| AS2 | 5′-CTCCTTGGGGtAcCACAATCAC-3′ | (−) KpnI |
| AS3 | 5′-catgtacccgggtGAACCTTCACAGAAAT TAAAGGC-3′ | (+) SmaI |
| AS4 | 5′-catgtacccgggCtATTGCCCTTTGACTT GTTGA-3′ | (−) SmaI |
| AS6 | 5′-CAGTAAAACCGgAtCcGACTTGG-3′ | (−) BamHI |
| SP1 | 5′-CGTTAATATCCAGGATCAAGGC-3′ | (−) RACE PCR |
| SP2 | 5′-GCTTCTCGTTTCTTTTTCCGCC-3′ | (−) RACE PCR |
| SP3 | 5′-GTCCTTTTTGGATGGCCTTC-3′ | (−) RACE PCR |
| Pgd5 (SP3) | 5′-TGATTCCTTTTCTAACTTGGTC-3′ | (−) RACE PCR |
| Pgd6 (SP2) | 5′-TAGGACTTGTGGGGTTATTTG-3′ | (−) RACE PCR |
| Pgd7 (SP1) | 5′-CCCGTTTGGATATAACGTTGC-3′ | (−) RACE PCR |
| 1934C (SP3) | 5′-CTAGCATACCATTAAAAAATGG-3′ | (−) RACE PCR |
| 1934D (SP2) | 5′-TCTTGATTTGTTGATAATCTGC-3′ | (−) RACE PCR |
| 1934E (SP1) | 5′-GCTTGTTAATTTTCAACATTTCG-3′ | (−) RACE PCR |
| 0159C (SP3) | 5′-GGCACTTAGGATAATTGCCAA-3′ | (−) RACE PCR |
| 0159D (SP2) | 5′-GCCATCAATCGAGCCACTAT-3′ | (−) RACE PCR |
| 0159E (SP1) | 5′-TGGCGTGAATAACGCACCAA-3′ | (−) RACE PCR |
| 0315C (SP3) | 5′-ATGCTCTAGAAAAGGAGCACT-3′ | (−) RACE PCR |
| 0315D (SP2) | 5′-TTCTACTTCTTTCGTCCACTC-3′ | (−) RACE PCR |
| 0315RT2 (SP1) | 5′-GCCAATTCCCGATAAAATCC-3′ | (−) RACE PCR |
| dC-Anchor | 5′-GACCACGCGTATCGATGTCGA C15D*-3′ | RACE PCR |
a
Nucleotides in lowercase are not complementary to the target sequence. Underlined nucleotides indicate sites inserted within primer sequences and corresponding to the restriction enzymes reported in the right column. (+), primer directed toward the 3′ end of the gene; (−), primer in the opposite direction. The indicated RACE PCR primers were used for first-strand cDNA synthesis (SPI), PCR amplification (SP2), or the sequencing reaction (SP3). *D = A, G or T.