Table 1. Bacterial strains, plasmids and primers used in this study.
| Strain | Description | Reference |
| XL1-Blue | supE44 hsdR17 recA1 endA1 gyrA46 thi relA1 lac−,[proAB+ lacIq lacZΔM15 Tn10 (TetR)] | Bullock et al., 1987 |
| BL21 pRep4-GroESL | F′, dmc, ompT, hsdS (r−Bm−B) galλ, pRep4-groESL | Amrein et al., 1995 |
| Plasmid | Description | Reference |
| pQE30 | Expression vector generating His6 fusion protein, AmpR | Qiagen |
| pGEXVM | Expression vector generating GST fusion protein, AmpR | Molle et al., 2003 |
| pQE30-Cap5A1B1 | Encoding last 29 amino acids of Cap5A1 (from Val194 to Asn222) fused to Cap5B1 (from Met1to Glu228), His6A1CtB1, cloned in BamHI/PstI sites, AmpR | Soulat et al., 2006 |
| pQE30-Cap5A1B2 | Encoding last 29 amino acids of Cap5A1 (from Val194 to Asn222) fused to Cap5B2 (Met1 toSer230), His6A1CtB2, cloned in BamHI/HindIII sites, AmpR | Soulat et al., 2006 |
| pQE30-Cap5O | Encoding Cap5O, His6CapO (from Met2 to Lys437), cloned in BamHI/PstI sites, AmpR | Soulat et al., 2007 |
| pQE30-Cap5A1B1 KKE/NTR | Same as pQE30-Cap5A1B1 but mutated on K3N,K4T and E5R | This study |
| pQE30-Cap5A1B2 NTR/KKE | Same as pQE30-Cap5A1B2 but mutated on N3K,T4K and R5E | This study |
| pGEXVM-A1Ct | Encoding last 29 amino acids of Cap5A1 from Val194 to Asn222, GSTA1Ct, cloned inBamHI/HindIII sites, AmpR | Soulat et al., 2006 |
| pGEXVM-A1CtΔN222 | Same as pGEXVM-A1Ct but deleted of Asn222 | This study |
| pGEXVM-A2Ct | Encoding last 27 amino acids of Cap5A2 from Leu194 to Phe220, GSTA2Ct, cloned inBamHI/HindIII sites, AmpR | Soulat et al., 2006 |
| pGEXVM-A2Ct+N222 | Same as pGEXVM-A2Ct but with a additional C-terminal Asn | This study |
| pET15b-A1-FL | Encoding Cap5A1 from Met1 to Asn222, His6A1, cloned in NdeI/BamHI sites, AmpR | Soulat et al., 2006 |
| pET15b-B1 | Encoding Cap5B1 from Met1 to Glu228, His6B1, cloned in NdeI/BamHI sites, AmpR | Soulat et al., 2006 |
| pET15b-B2 | Encoding Cap5B1 from Met1 to Ser230, His6B2, cloned in XhoI/BamHI sites, AmpR | Soulat et al., 2006 |
| Primer | 5′ to 3′ sequence a,b,c | Reference |
| B1 (−) | TGCACTGCAGTTATTCATCTCCATAATAGTGAT | This study |
| B2 (−) | CCCAAGCTTTCATGATTCATCAGTCCCATAATA | This study |
| A1ct (+) | TATGGATCCAAAGTAATTTTCGATAAGCG | This study |
| A2ct (+) | TATGGATCCTTATTAGATAAGCGTATTAAGACAG | This study |
| A1B2 N3KT4K R5E (−) | CTTGATGTACTTCTTCTTTCTTTTTTCGTCATATTAAATTTTTG | This study |
| A1B1 K3NK4T E5R (−) | GTGTTGTTGTCGTATTTCGCGTATTTGACATATTAAATTTTTG | This study |
| A1ctΔN222 (−) | TATAAGCTTTTAAAATTTTTGAATTGAACCCAATAC | This study |
| A2ct+N222 (−) | TATAAGCTTTTAATTAAATTTTTGTATTGAACCTAAAATAGG | This study |
a
Forward and reverse primers are represented by plus (+) or minus (−), respectively.
b
restriction sites are italicized.
c
The bases mutated from those present in the wild type are bold.