Table 2.
Plasmid | Primer or fragment used* | Construction† |
---|---|---|
pLJH1 (rsbT1) | T-1 5′ TGCGGATCCATTCCAGGAAGATTTGC 3′ | pKSV7 BamHI–PstI |
T-2 5′ GGACCTTTGTTTTTGGCAAC 3′ | ||
T-3 5′ GTTGCCAAAAACAAAGGTCC 3′ | ||
T-4 5′ TTCCTGCAGAGCAATTTCACTACGTA 3′ | ||
pLJH2 (rsbUΔ1) | U-1 5′ CTCGGATCCACGTTTAGCGGAGTAC 3′ | pKSV7 BamHI–PstI |
U-2 5′ CACTTCTTCTTATTTAGTTTTTTCCATTTTTTTCTAGCT 3′ | ||
U-3 5′ ACTAAATAAGAAGAAGTGAAG 3′ | ||
U-4 5′ AGGCTGCAGAGGAAGTTGTAATGTAA 3′ | ||
pLJH3 | F 5′ CTTCCTGCAGAATGTAACACTGTCTTT 3′ | pPL2 PstI–BamHI |
R 5′ TTTGGATCCTTTATAAGTCTTCGTTAC 3′ | ||
pLJH4 | BglII–BamHI from pHK77, encoding spoVG–lacZ fusion | pLJH3 SacI–BamHI |
pLJH5 (rsbRA) | F 5′ GAAAGGATCCGATGTATAAAGATTTTGCA 3′ | pET-15b BamHI–BlpI |
R 5′ CCCGCTCAGCTCACCCCTCTTTTTCTAC 3′ | ||
pLJH6 (rsbS) | F 5′ GGTGGATCCTGTGGGGATACCAATCTTA 3′ | pET-15b BamHI–BlpI |
R 5′ GAATGCTCAGCTCATTCCCCCAATTCCTG 3′ | ||
pLJH7 (rsbT) | F 5′ GGGGGATCCCATGACATTCCAATCCTGT 3′ | pET-15b BamHI–BlpI |
R 5′ CTAGCTCAGCTTACCGAACCCATTTCGT 3′ |
*Restriction endonuclease sites used for cloning are denoted by underlined italic type.
†Vector and sites used: pKSV7 (Smith & Youngman, 1992); pPL2 (Lauer et al., 2002); pET15b (Novagen).