Table 2.
Primers and PCR products used in this study.
| Primers, PCR products, or bacteriocins | Nucleotide sequence (5′-3′) or description | Amplifications |
|---|---|---|
| Primers | ||
| S602-F | GCCATGAGCTCGAATTCTCGAGAAAAG | R-SRCAM602, R-E760, R-L1077 |
| S071-F | GTCCAGAGCTCGAATTCTCGAGAAAAG | R-SRCAM 602, R-E760, R-L1077, R-OR7 |
| SARP-R | AGGTACCATAAGTTGCGGCCGC | R-OR7 |
| ALFA-F | TACTATTGCCAGCATTGCTGC | pPICZαA amplification fragment including the cloned synthetic gene |
| 3AOX1-R | GCAAATGGCATTCTGACATCC | pPICZαA amplification fragment including the cloned synthetic gene |
| PCR products | ||
| R-SRCAM602 | 136-bp XhoI/NotI fragment containing the α-factor Kex2 signal cleavage fused to the mature synthetic srcam602 gene with the P. pastoris codon usage | |
| R-OR7 | 181-bp XhoI/NotI fragment containing the α-factor Kex2 signal cleavage fused to the mature synthetic or-7 gene with the P. pastoris codon usage | |
| R-E760 | 242-bp XhoI/NotI fragment containing the α-factor Kex2 signal cleavage fused to the mature synthetic e-760 gene with the P. pastoris codon usage | |
| R-L1077 | 167-bp XhoI/NotI fragment containing the α-factor Kex2 signal cleavage fused to the mature synthetic l-1077 gene with the P. pastoris codon usage | |
| Bacteriocins | ||
| BacSRCAM602 (amino acid sequence) | ATYYGNGLYCNKQKHYTWVDWNKASR EIGKITVNGWVQH |
|
| BacSRCAM602 (P. pastoris codon usage) | gctacttactacggtaacggtctttactgtaacaagcagaagcact acacttgggttgactggaacaaggcttccagagagatcggtaag atcactgttaacggttgggttcaaca |
|
| BacOR-7 (amino acid sequence) | KTYYGTNGVHCTKNSLWGKVRLKNMK YDQNTTYMGRLQDILLGWATGAFGKTFH |
|
| BacOR-7 (P. pastoris codon usage) | aagacttactacggaactaacggtgttcactgtactaagaattcctt gtggggtaaggttagattgaagaacatgaagtacgaccagaaca ctacttacatgggtagattgcaggacatcttgttgggttgggctact ggtgctttcggtaagacttttcat |
|
| BacE-760 (amino acid sequence) | NRWYCNSAAGGVGGAAVCGLAGYVGE AKENIAGEVRKGWGMAGGFTHNKACKS FPGSGWASG |
|
| BacE-760 (P. pastoris codon usage) | aacagatggtactgtaactccgctgctggtggtgttggtggtgct gctgtttgtggtttggctggttatgttggtgaggctaaagaaaacat tgctggtgaggttagaaagggttggggtatggctggtggtttcac tcataacaaggcttgtaagtccttcccaggttctggttgggcttctggt |
|
| BacL-1077 (amino acid sequence) | TNYGNGVGVPDAIMAGIIKLIFIFNIRQGY NFGKKAT |
|
| BacL-1077 (P. pastoris codon usage) | actaactacggtaacggtgttggtgttccagacgctattatggctg gtatcatcaagttgatcttcatcttcaacatcagacagggttacaac ttcggtaagaaggctact |