Table 2. Strains, plasmids, and primers used in this study.
| Strain or plasmid | Characteristic or relevant feature | Reference or source |
| E. coli | ||
| DH5α | F- φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (rk−, mk+) phoA supE44 λ- thi-1 gyrA96 relA1 | Invitrogen |
| BL21 | F- ompT hsdSB (r B −m B −) gal dcm rne131 (DE3) | Invitrogen |
| DB3.1 | F- gyrA462 endA1 glnV44 Δ(sr1-recA) mcrB mrr hsdS20(rB −, mB −) ara14 galK2 lacY1 proA2 rpsL20(Smr) xyl5 | Invitrogen |
| XL1-Blue | recA1 endA1 gyrA96 thi-1 hsdR17 (rk −, mk −) supE44 relA1 lac (F− proAB lacI q Z ΔM15 Tn10 (tetr)) | Stratagene |
| Plasmids | ||
| pDONR221 | Entry vector, Kmr | Invitrogen |
| pDEST17 | Destination expression vector of the Gateway system (His-tag), Apr | Invitrogen |
| pDEST24 | Destination expression vector of the Gateway system (GST), GST donor, Apr | Invitrogen |
| pTP145 | Phage display vector, Cmr | 37 |
| pKRP13 | Spectinomycin resistance gene donor, Spr | 53 |
| pTP245 | Phage display vector derived from pTP145, containing the attR1-Cmr-ccdB-attR2 sequence, Apr | This study |
| pTP247 | Expression vector derived from pDEST17, containing a stop codon at the position 1846, Apr | This study |
| pTP248 | Expression vector derived from pDEST17, containing both His- and GST tags, Apr | This study |
| pTP249 | Expression vector derived from pDEST17, containing a DNA fragment encoding a pelB signal sequence, Apr | This study |
| pTP262 | Phage display vector derived from pTP145, containing the attR1-Sper-ccdB-attR2 sequence, Cmr, Spr | This study |
| Primers | ||
| attB1 forward | GGGGACAAGTTTGTACAAAAAAGCAGGCTTC-ORF specific bps1 | Invitrogen |
| attB2 reverse | GGGGACCACTTTGTACAAGAAAGCTGGGTN-ORF specific bps2 | Invitrogen |
| DONRF-2 | GTTCCCTACTCTCGCGTTAACG | This study |
| DEST17-STOP | CAGCTTTCTTGTACAAAGTGGTTTGATTCGAGG | This study |
| PELB-F | TATGAAATACCTGCTGCCGACCGCTGCTGCTGGTCTGCTGCTCCTCGCTGCCCAGCCGGCGATGGCCCA | This study |
| PELB-R | ACTTTATGGACGACGGCTGGCGACGACGACCAGACGACGAGGAGCGACGGGTCGGCCGCTACCGGGTAT | This study |
| DEST-XHOI | GGGCTCGAGCTCTCGAATCAACAAGTTTGTACAAAAAAGCTGAACGAGAAACGTAAAATG | This study |
| DEST-XBAI | GGCGTCTAGACAACCACTTTGTACAAGAAAGCTGAACGAGAAACGTAAAATGATATAAATATC | This study |
| NPAP-F | TTGCGAGGCTCAGTTTTATCAC | This study |
| NPAP-R | CCAGAATTGGCGTAAAACATCA | This study |
1,
The 25 bp attB1site is underlined.
2,
The 25 bp attB2 site is underlined, N represents A, G, C, or T.