Table 1.
Oligonucleotide primer sequences.
| Purpose | Primer sequence |
|---|---|
| Reverse transcriptase—PCR: nitr9 | GGATTTTTGGACTTTTCTGTC |
| TCCACATGCGGTAACTGTAC | |
| Reverse transcriptase—PCR: β-actin | GGTATGGAATCTTGCGGTATCCAC |
| ATGGGCCAGACTCATCGTACTCCT | |
| TaqMan Q-PCR: nitr9 (probe = CAAGGTTTGGAAAAGCAC) | GTCAAAGGGACAAGGCTGATAGTT |
| GTTCAAAACAGTGCATGTAAGACTCA | |
| TaqMan Q-PCR: β-actin (probe = CCCATGCCATCCTGC) | CCATCTATGAGGGTTACGCTCTTC |
| AGGATCTTCATCAGGTAGTCTGTCA | |
| Amplify nitr9 I domain for bacterial expression construct | A TGGAAAAGCACACTGTAGTAa |
| TTATTTAGAGCCATTCCTGTCCb | |
| Amplify nitr9L for FLAG-tagged expression cassette | CACCCAAATGCACCACCTGTGTTTGTTAAACc |
| gactgcggccgcTTACTGCTGGTTAGAAACd | |
| Amplify nitr9S for FLAG-tagged expression cassette | CACCCAAATGCACCACCTGTGc |
| gactgcggccgcTTACTGCTGGTTAGAAACd | |
| Amplify nitr9SS for FLAG-tagged expression cassette | CATGATTTAATTCCATCCCAc |
| gactgcggccgcTTACTGCTGGTTAGAAACd | |
| Amplify wild type nitr9L, nitr9S and nitr9SS for expression cassettes | gatcggatccgacATGATCAACTTTTGGATTTe |
| gatcgaattcTTACTGCTGGTTAGAAACCGAGf |
aAn artificial start codon is underlined.
bAn artificial stop codon is bold.
cThese primers are designed for blunt PCR cloning into the EcoRV site of pLF.
dOverhang (5′) sequences are in lower case text and include a Not I site for cloning into pLF.
eOverhang (5′) sequences are in lower case text and include a BamHI site for cloning into pcDNA3.
fOverhang (3′) sequences are in lower case text and include an EcoRI site for cloning into pcDNA3.