TABLE 1.
Strains, plasmids, and primers used in this study
Restriction sites are denoted in lowercase type.
| Name | Description | Reference/Source |
|---|---|---|
| Plasmids | ||
| pKS505 | Gurken TMD in pKS29; ampr | Ref. 8 |
| pKS506 | LacY-TM2 in pKS29; ampr | Ref. 8 |
| pKS507 | Dm Spitz TMD in pKS29; ampr | Ref. 8 |
| pKS508 | PstI TatA aa1–50 into pKS29; ampr | Ref. 8 |
| pTA131 | Ampr; pBluescript II containing Pfdx-pyrE2 | Ref. 26 |
| pMIG1 | pTA131 containing rhoII-flanking regions | This study |
| pTA963 | AmpR, overexpression vector with His6 tag, pyrE2 and hdrB markers, and pHV2 origin. | Ref. 45 |
| pMCF1 | pTA963 with region 207 bp upstream of Hvo_0727 start codon and 103 bp downstream of Hvo_0726 stop codon, cloned between the ApaI and BamHI sites. | This study |
| Strains | ||
| E. coli DH5α | E. coli F− φ80dlacZΔM15 (lacZYA-argF)U169 recA1 endA1 hsdR17 (rK− mK+) phoA supE44 thi-1 gyrA96 relA1 | Invitrogen |
| E. coli GM33 | LAM-, IN(rrnD-rrnE)1, F− dam-3 sup-85 (Am) | Ref. 46 |
| E. coli Rossetta (DE3) | F_ ompT [Ion] hsdSB (rB mB) (an E. coli B strain) with DE3, a prophage carrying the T7 RNA polymerase gene | Novagen |
| E. coli MG1655 | ΔglpEGR::kan | |
| H. volcanii H26 | H. volcanii DS70 ΔpyrE2 | Ref. 28 |
| H. volcanii MIG1 | H26 ΔrhoII | This study |
| Primers | ||
| HindFupRho2 | 5′-AGaagcttTTCATCCCGCAGACGT-3′ | This study |
| RevUpsRho2ecoRI | 5′-TgaattcATTACCAACGCCTTA-3′ | This study |
| BamFdownRho2 | 5′-AggatccGAGATGCGGCCGGCCCGTC-3′ | This study |
| RevdownRHO2Xba | 5′-AtctagaTCTCAGTCTTCGTACT-3′ | This study |
| Hvo0727KOverify5′ | 5′-TACGAGTACGACAAGGACAT-3′ | This study |
| Hvo0727KOverify3′ | 5′-TAAGTGTACGCGGTTCGCCA-3′ | This study |
| 7 S F | 5′-CCAACGTGGAAGCCTCGTC-3′ | Ref. 21 |
| 7 S R | 5′-GGTGGTCCGCTGCTCACTTC-3′ | Ref. 21 |
| FwOperonRhoII | 5′-GTGATGAGGACCGTCTTTTG-3′ | This study |
| RvOperonRhoII | 5′-CGCGTAATCTCGCATTTCTC-3′ | This study |