Table 1.
Primers designed to amplify S. aureus Newman strain ORFs for protein expression
| Gene locus no. | Function | Forward primer (5′ to 3′)a | Reverse primer (5′ to 3′)b |
|---|---|---|---|
| NWMN_0161 | Putative lipoprotein | ATGGGAAAATCAAAAATTTATATCTTG | TGAATCATCTCCAAAAATTTATG |
| NWMN_0364 | Putative lipoprotein | ATGGGAAAATTAAAATCATTAGCAG | GTGATCTTGCTCACTCTTTAATAC |
| NWMN_0389 | Enterotoxin | ATGGGAAAAATGAAAAATATTGC | TGCTTTTATAACTTTGATTTC |
| NWMN_0390 | Exotoxin 8 | ATGGGAAAAATGAGAACAATTGC | CTTATCATTAGCATTATTTTG |
| NWMN_0429 | Alanine amidase | ATGGGAGTGCAAAAAAAAGTAATTGCAGC | GTGAATATATCTATAATTATTTAC |
| NWMN_0458 | Erythritol kinase | ATGGGAATATATGAAACGGCACCAG | TCCTAATAGTCTAACTAAGTAC |
| NWMN_0601 | Putative lipoprotein | ATGGGAAAAAAATTAGTACCTTTA | TTCATGCTTCCGTGTACAGTTTC |
| NWMN_0668 | Fructose PO4 kinase | ATGGGAATTTATACAGTGACTTTC | CTCCCCATCAAGTACGCTAAT |
| NWMN_0724 | LysM domain protein | ATGGGAAAAAAATCTCTTACAGTG | GTGGATGTAATTATATTTTCCTG |
| NWMN_0734 | CofD-like proteins | ATGGGAAGACAAATAAAAGTTGTAC | TTTACGTTTATCACTTGGTACG |
| NWMN_0738 | Putative protein | ATGGGATCGAATCAAAATTACG | GTTATCATTAGCATTATTTTG |
| NWMN_0753 | Putative lipoprotein | ATGGGAAAAAAAGTAATGGGGATA | TACTTCATCTAAACCACTGTGG |
| NWMN_0791 | CBS domain protein | ATGGGAGTGATCATTGCCATAATTA | TATAGATACGTTGCGATTTTTCCG |
| NWMN_0925 | LytR domain protein | ATGGGAAATAAATTTTTAAAATAC | ATTTACAACATTTTGG |
| NWMN_1067 | FPRL1 inhibitory protein | ATGGGAAAAAAAAATATCACAAAAAC | ATACCAAGTAATCGAGTCGATTTC |
| NWMN_1069 | Fibrinogen-binding protein | ATGGGAAAAAATAAATTGATAGC | TTTAACTAATCCTTGTTTTAATAC |
| NWMN_1236 | Thermonuclease | ATGGGAAAGTCAAATAAATCGCTTG | TTTACTCCAAATATTTAATTTC |
| NWMN_1398 | Putative lipoprotein | ATGGGAAAAAAATTCATTGGATCAG | TTCATCTTTACTGTGCACCCCATATTC |
| NWMN_1607 | Thiol peroxidase | ATGGGAACTGAAATAACATTCAAAG | AATATTTTTGTATGCAGCTAAAGC |
| NWMN_1623 | Glycosyl transferase | ATGGGAACGAATCAAGACAATC | ACGCTTTTTTCGATTTAACAAATG |
| NWMN_1690 | Putative lipoprotein | ATGGGAAAATTCAAAGCTATCGTTG | TTTAACTTTAGTTTCTTCAG |
| NWMN_1701 | Serine protease SplF | ATGGGAAATAAAAATATAATCATCAAAAGTATTGCAG | TTTATCTAAATTATCTGCAATG |
| NWMN_1703 | Serine protease SplD | ATGGGAAATAAAAATATAATCATCAAAAGTATTGCG | TTTATCTAAATTATCTGCAATAAATTTC |
| NWMN_1704 | Serine protease SplC | ATGGGAAATAAAAATATAGTCATTAAAAGC | TTGTTCAATGTGCTTTTGAATAAAATC |
| NWMN_1705 | Serine protease SplB | ATGGGAAACAAAAACGTAGTCATCAAG | TTTATCTATGTTTTCTGCAATG |
| NWMN_1706 | Serine protease SplA | ATGGGAAATAAAAATGTAATGGTTAAAG | TTTTTCAATATTATTTTGAATAAATTG |
| NWMN_1726 | RNAIII-activating protein | ATGGGAAAGAAACTATATACATC | TTTCATGCTTCCGTGTACAGTTTC |
| NWMN_1733 | Foldase protein | ATGGGAAAGATGATAAACAAATTAATC | TTGGCTCATGCCGGATTGTC |
| NWMN_1876 | Complement inhibitor | ATGGGAAAAATTAGAAAATCTATAC | ATATTTACTTTTTAGTGCTTC |
| NWMN_1880 | Staphylokinase | ATGGGACTCAAAAGAGGTTTATTA | TTTCTTTTCTATAATAACCTTTG |
| NWMN_1926 | β-Hemolysin | ATGGGATATCCAAACTGGGGGC | TTTACTATAGGCTTTGATTGG |
| NWMN_1999 | Transglycosylase sceD | ATGGGAAAGAAAACATTACTCGCATC | TGCAGTAACCCAATGTCCAGC |
| NWMN_2043 | Dps family protein | ATGGGAAGTAATCAACAAGATGTTG | GCTTAAGTAAGATTTAAACATC |
| NWMN_2086 | Alkaline shock protein | ATGGGAACTGTAGATAACAATAAAG | TTGTAAACCTTGTCTTTCTTGG |
| NWMN_2270 | Putative lipoprotein | ATGGGAAAAAGATTAGTTACAGGGTTAC | TTGTTGGTAGTTTGGATCAGTAAC |
| NWMN_2318 | γ-Hemolysin A | ATGGGAATTAAAAATAAAATATTAAC | CTTAGGTGTGATGCTTTTAATTTTTAC |
| NWMN_2319 | γ-Hemolysin C | ATGGGACTTAAAAATAAAATATTAAC | ATTCTGTCCTTTCACCTTGATTTC |
| NWMN_2320 | γ-Hemolysin B | ATGGGAAAAATGAATAAATTAGTC | TTTATTGTTTTCAGTTTCTTTTG |
| NWMN_2426 | Sortase A | ATGGGAAAAAATGGACAAATCG | TTTGACTTCTGTAGCTACAAAG |
| NWMN_2469 | Antigen A | ATGGGAAAAAAGACAATTATGGCATC | GAATCCCCAAGCACCTAAACCTTG |
| NWMN_2537 | Antigen B | ATGGGAAATAAAACCAGTAAAGTTTG | TTTACTTGTTTTGTATGGTGTATG |
| NWMN_2544 | Isochorismatase | ATGGGATCTCGAAAAACGGCGCTATTAG | TAAACTACTTTTCCATGACTC |
| NWMN_2579 | Putative lipoprotein | ATGGGACATAAAAAATATTTATTGC | TCTTCATAATCCCGATTCCC |
a
The forward primers were designed with a CACC overhang at the 5′ end to facilitate directional cloning.
b
The reverse primers were designed with a TCATTTGTCGTCGTCGTCTTTATAGTC overhang to incorporate a FLAG epitope and a stop codon at the 3′ end.