TABLE 1.
Materials used for the expression of the different Saptp proteins in P. pastoris and for RT-PCR
| Genes | Oligonucleotide primer (sequence) | Orientation | Encoded amino acid sequencea | Template | PCR product (with cloning sites) |
|---|---|---|---|---|---|
| SAPT1 | 1 (GGA A/GA TCT GAT GTG CCA ACT ACA TTG A) | Senseb | pMTL21-E4 or cDNA | SAPT1 (181–1,186 bp); BglII----NotI | |
| 2 (CGT GC/GGC CGC T CTA CAA AGC CGA GAT GTC T) | Antisenseb | ||||
| 3 (GTA C/TC GAG AAA AGA TTG TTT GCT CAA GGT CTT ACT) | Sensec | (L)(E)(K)(R)LFAQGLT | pMTL21-E4 | SAPT1 (55–1,185 bp); XhoI----NotI | |
| 4 (CAT GC/GC CGC CTA CAA AGC CGA GAT GTC TGA AGA) | Antisensec | SSDISALSTOP | |||
| SAPT2 | 5 (GTA C/T CGA GTT TCC AAG CTC AAC GAT CGT) | Sensec | (R)(V)SKLNDR | pCT2 | SAPT2 (55–511 bp); XhoI----Asp718I |
| 6 (C TTT GAC CCA G/GT ACC ACT AGA AGA A) | Antisensec | SSSGTWVK | |||
| 7 (T TCT TCT AGT G/GT ACC TGG GTC AAA G) | Sensebc | SSSGTWVK | pCT2 or cDNA | SAPT2 (486–1,248 bp); Asp718I----BglII | |
| 8 (CAT A/GA TCT CTA AAC AAT AGT GAC ATT AGA) | Antisensebc | SNVTIVSTOP | |||
| SAPT3 | 9 (GTA C/T CGA GTG GTT TTA AAT GCT GGT TCA) | Sensebc | (R)(V)VLNAGS | pCT3 or cDNA | SAPT3 (52–1,170 bp); XhoI----BglII |
| 10 (CAT A/GA TCT CTA CAT AAT TGC TCT AAT TTT) | Antisensebc | KIRAIMSTOP | |||
| SAPT4 | 11 (GTA C/T CGA GCT CCT ACA ACT TCA CCT CCT) | Sensebc | (R)(A)PTTSPP | pCT4 or cDNA | SAPT4 (55–1,185 bp); XhoI---- BamHI |
| 12 (CAT G/GA TCC CTA TGT AAG TGG AAG TAT GTT) | Antisensebc | NILPLTSTOP |
a
In parentheses are shown amino acids encoded by the XhoI restriction site sequence and added to the N-terminal extremity of the Saptp prosequences.
b
Primers used for RT-PCR.
c
Primers used for expression in P. pastoris.