Table 2.
Plasmids.
| Plasmid | Relevant properties | Derivation |
|---|---|---|
| pGB70 | Integrative plasmid based on pUC19, with pyrE2 marker | [36] |
| pTA50 | pBluescript II with Eco47III-XmaI chromosomal fragment containing radB | [16] |
| pTA62 | pGB70 with radBΔb/b partial deletion, generated by excision of BstBI-BstEII fragment of radB from pTA50 | This study |
| pTA131 | Integrative plasmid based on pBluescript II, with pyrE2 marker | [18] |
| pTA163 | Integrative plasmid containing leuB-Aa2 allele, for use in recombination assay | [20] |
| pTA289 | pTA131 with ΔradB construct | [16] |
| pTA311 | pTA131 with radB+, generated by insertion of KpnI-BspEI fragment of pTA50 containing radB | This study |
| pTA324 | pTA131 with ΔradA:trpA+ construct | [25] |
| pTA409 | Shuttle vector based on pBluescript II, with pyrE2 and hdrB markers and ori-pHV1 origin | [25] |
| pTA411 | pTA409 with radA+ gene, for complementation of ΔradA | [25] |
| pTA769 | pTA131 with radA-A196V, generated by PCR of KpnI-BstBI radA-A196V fragment from H188 | This study |
| pTA963 | Overexpression vector with p.tnaA promoter, 6xHis tag, pyrE2 and hdrB markers, and pHV2 origin | [19] |
| pTA1043 | pTA963 with radB, for overexpression of 6xHis-tagged RadB | [19] |
| pTA1289 | pTA131 with radA-S101P, generated from pTA769 by replacement with AgeI-BstEII radA-S101P fragment from H1309 | This study |
| pTA1314 | pTA131 with radA-S101P-A196V, generated from pTA1289 by replacement with AflIII fragment from pTA769 | This study |
| pTA1539 | pTA131 with ΔradB:trpA+ construct, generated by PCR of XhoI-BamHI fragment of upstream flanking region and BamHI-XbaI fragment of downstream flanking region from pTA50, with insertion of trpA+ BamHI fragment of pTA298 [20] | This study |
| pTA1771 | pTA131 with insertion of his7tag-2xStrepIItag cassette at EcoRV site in multiple cloning site, features NdeI site upstream of 7xHis tag and PciI site downstream of 2xStrepII tag | This study |
| pTA1815 | pTA1771 with insertion of FatI-BamHI radB+ fragment of pTA1043, at PciI and BamHI sites | This study |
| pTA1847 | pTA1539 with replacement of ΔradB:trpA+ by NdeI-BamHI fragment of pTA1815 with his7tag-2xStrepIItag-radB+ allele | This study |
| pTA1868 | pTA131 with radA-S101A, generated from pTA1289 by replacement with AgeI-BstEII PCR fragment using radAS101Aint primers | This study |