TABLE 2.
Plasmids and oligonucleotides used in this study
| Plasmid or oligonucleotide | Descriptiona | Reference |
|---|---|---|
| General purpose vectors | ||
| pBKS+/− | General cloning vector (pBKS); Ampr | 54 |
| pBSK− | General cloning vector (pBSK); Ampr | 54 |
| pRKlac290 | lacZ transcriptional fusion; Tetr | 1 |
| pUJ142 | Xylose-inducible promoter; Chlr | 30 |
| Subclones of the podJ region | ||
| pBC1 | 151-bp PCR fragment (−128 to +24) generated with primers | This study |
| pDZ7 | 3-kb PstI subclone from K28 containing podJ in pBKS− | This study |
| pBC3 | 860-bp PstI/XhoI fragment of pDZ7 in pBSK+ | This study |
| pBC4 | 1,519-bp PstI/HindIII fragment of pDZ7 in pBSK+ | This study |
| pBC5 | 1,519-bp PstI/HindIII fragment of pDZ7 in pBKS+ | This study |
| pBC9 | 95-bp fragment (−128 to −33) in pBKS+ from the StyI/XbaI deletion of pBC1 | This study |
| pBC12 | 211-bp PCR fragment (−188 to +24) generated with primers BE364 and BE238, in HindIII site of pBKS+ | This study |
| pBC15 | 201-bp fragment (−188 to −148, −137 to +24) resulting from a HpaI deletion of pBC12 | This study |
| Expression fusion | ||
| pBC16 | 1,614-bp PCR fragment generated with primers BE300 and BE301, cloned into the SpeI/KpnI sites of pUJ142 (PxylX::podJ) | This study |
| Transcriptional fusions | ||
| pBC6 | pBC5 into the HindIII/KpnI sites of pRKlac290 | This study |
| pBC7 | pBC4 into the PstI/XbaI sites of pRKlac290 | This study |
| pBC10 | pBC1 into the HindIII/KpnI sites of pRKlac290 | This study |
| pBC11 | pBC9 into the HindIII/KpnI sites of pRKlac290 | This study |
| pBC13 | pBC12 into the KpnI site of pRKlac290 | This study |
| pBC17 | pBC15 into the KpnI site of pRKlac290 | This study |
| Oligonucleotides | ||
| BE176 | 5′-ACCAACCTCATCTGCCGGAT | This study |
| BE208 | 5′-AACTGCAGGCTTTTGAACCGCGCTACAGAG | This study |
| BE238 | 5′-CCCAAGCTTCGTCATGCGAATCGATCTCC | This study |
| BE364 | 5′-CCCAAGCTTGCGCGCTTCTAGGCCTGCGACA | This study |
| BE369 | 5′-CTAGGCGGCCAGATCTGATC | This study |
a
Antibiotic resistance markers: Ampr, ampicillin; Tetr, tetracycline; Chlr, chloramphenicol. Nucleotide positions given in parentheses are in relation to the podJ transcription start site. pBKS, pBluescript KS; pBSK, pBluescript SK.