Table 1.

Materials used for the expression of the different Saptp proteins in P. pastoris and for RT-PCR

GenesOligonucleotide primer (sequence)OrientationEncoded amino acid sequenceaTemplatePCR 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) Senseb,c SSSGTWVK pCT2 or cDNA SAPT2 (486–1,248 bp);Asp718I----BglII
8 (CAT A/GA TCT CTA AAC AAT AGT GAC ATT AGA) Antisenseb,c SNVTIVSTOP
SAPT3 9 (GTA C/T CGA GTG GTT TTA AAT GCT GGT TCA) Senseb,c (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) Antisenseb,c KIRAIMSTOP
SAPT4 11 (GTA C/T CGA GCT CCT ACA ACT TCA CCT CCT) Senseb,c (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) Antisenseb,c 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.