TABLE 1.
Strains, plasmids, and primers used in this study
| Strain, plasmid, or primer | Properties or 5′-to-3′ primer | Reference or source |
|---|---|---|
| Strains | ||
| R. capsulatus SB1003 | Wild type, Rifr | |
| R. capsulatus SB1003 ΔhupSL | ΔhupSL, Rifr | This study |
| R. capsulatus SB1003 ΔRRC00247-RRC00248 | ΔRRC00247-RRC00248, Rifr | This study |
| E. coli UQ950 | Cloning strain | D. Liesb |
| E. coli WM3064 | Donor strain | W. Metcalfc |
| S. cerevisiae InvSC1 | Ura− for gap repair cloning | Invitrogen |
| Plasmids | ||
| pMQ30 | Yeast-based allelic exchange vector, sacB,a CEN/ARSH, URA3+, Gmr | 35 |
| pMQ87 | Yeast-based suicide vector, CEN/ARSH, URA3+, Gmr | 35 |
| pZJD29a | Allelic exchange vector, sacB,a Gmr | C. Bauerd |
| pMQ131 | Yeast-based pBBR1-based shuttle vector, CEN/ARSH, URA3+, Knr- | R. M. Q. Shanks and G. A. O'Toolee |
| pNC001 | hupSL deletion fragments cloned into pMQ30 | This study |
| pNC005 | hupSL deletion fragments cloned into pZJD29a | This study |
| pNC006 | Yeast-based allelic exchange vector, sacB,a CEN/ARSH, URA3+, Gmr | This study |
| pNC007 | RRC00247 and RRC00248 deletion fragment cloned into pNC006 | This study |
| pNC008 | RRC00247 and RRC00248 genes clones into pMQ131, Knr | This study |
| Primers | ||
| pnc001 | GTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTCTGGGCGAGAACCTTTGG | |
| pnc002 | CCTTGACGGTGGTCAGGCAATTGTCCCTCCCTTGC | |
| pnc003 | GCAAGGGAGGGACAATTGCCTGACCACCGTCAAGG | |
| pnc004 | GGCAAATTCTGTTTTATCAGACCGCTTCTGCGTTCT GATCTCTTCCAGAAACGGCAGC | |
| pnc017 | GGCGGAGCTCCTGGGCGAGAACCTTTGG | |
| pnc018 | GGCGTCTAGACTCTTCCAGAAACGGCAGC | |
| pnc021 | ATGCCACGATCCTCGCCCTGCTGGCGAAGATCGACTCTAGCCACAGTCGATGAATCCAG | |
| pnc022 | ATCTCTAAGAAACCATTATTATCATGACATTAACCCGACAATTCGACCTGAAAATTCC | |
| pnc023 | GGCAAATTCTGTTTTATCAGACCGCTTCTGCGTTCT GATGAAGATCGACGAGGTTCTGG | |
| pnc027 | AGGTTCGAGATGATCCGGTTGAAACCCTCCCGTTACCTTG | |
| pnc028 | CAAGGTAACGGGAGGGTTTCAACCGGATCATCTCGAACCT | |
| pnc026 | TGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTCGAGACGGCT TTCCACAGG | |
| pnc033 | GGCGTCTAGAATGTGAAGCACAACAGCACCTACG | |
| pnc034 | GGCGGAGCTCATAATGGCGCAGGTTCTGCCAAAG |
a
sacB in pMQ30 is under the control of the native promoter, and the sacB gene in pZJD29a and pNC006 is under the control of the R. capsulatus pucAB promoter.
b
California Institute of Technology, Pasadena, CA.
c
University of Illinois, Urbana, IL.
d
University of Indiana, Bloomington, IN.
e
Dartmouth College (www.dartmouth.edu/∼gotoole/vectors.html), Hanover, NH.