Table 1.
Bacterial strains and plasmids used in this study
| Strain or plasmid | Relevant characteristics and/or derivationa |
|---|---|
| Fox− derivatives of Anabaena sp. strain PCC 7120 | |
| FQ202 | Bmr Nmr Smr; alr1728::Tn5-1063 |
| FQ204 | Bmr Nmr Smr; all1591::Tn5-1063 |
| FQ211 | Bmr Nmr Smr; all3520::Tn5-1063 |
| FQ228 | Bmr Nmr Smr; all3278::Tn5-1063 |
| FQ324 | Bmr Nmr Smr; alr1728::Tn5-1063 |
| FQ384 | Bmr Nmr Smr; all3850::Tn5-1063 |
| FQ406 | Bmr Nmr Smr; all4019::Tn5-1063 |
| FQ747 | Bmr Nmr Smr; all1591::Tn5-1063 |
| FQ807 | Bmr Nmr Smr; alr4311::Tn5-1063 |
| FQ1265 | Bmr Nmr Smr; all1338::Tn5-1063 |
| FQ1470 | Bmr Nmr Smr; all1338::Tn5-1063 |
| FQ1580 | Bmr Nmr Smr; all3582::Tn5-1063 |
| FQ1595 | Bmr Nmr Smr; all4388::Tn5-1063 |
| SR2816a | Cmr Emr; mutation in all3850 resulting from single recombination with pRL2816a; see below |
| SR2821 | Cmr Emr; mutation in alr1728 resulting from single recombination with pRL2821; see below |
| Escherichia coli strains | |
| DH5αMCR | Invitrogen Corp., received courtesy of J. C. Meeks |
| DH10B | F−mcrA Δ(mrr-hsdRMS-mcrBC) φ80lacZΔM15 ΔlacX74recA1endA1araD139 Δ(ara leu)7697galUgalKrpsLnupG λ− |
| HB101 | F−thi-1hsdS20 (rB− mB−) supE44recA13ara-14leuB6proA2lacY1galK2rpsL20 (Smr) xyl-5mtl-1 |
| Plasmids | |
| anc0756 | Cmr Emr; Anabaena sp. chromosomal DNA from bp 4233705 to 4250432 in the BamHI site of pRL838 |
| anc1006 | Cmr Emr; Anabaena sp. chromosomal DNA from bp 3952462 to 3970608 in the BamHI site of pRL838 |
| anc1512 | Cmr Emr; Anabaena sp. chromosomal DNA from bp 2070875 to 2084189 in the BamHI site of pRL838 |
| anp00427 | Apr; Anabaena sp. chromosomal DNA from bp 5161083 to 5168600 in the BamHI site of pUC18 |
| anp00676 | Apr; Anabaena sp. chromosomal DNA from bp 4325494 to 4333569 in the BamHI site of pUC18 |
| anp00920 | Apr; Anabaena sp. chromosomal DNA from bp 4640078 to 4648267 in the BamHI site of pUC18 |
| anp02032 | Apr; Anabaena sp. chromosomal DNA from bp 5253901 to 5262374 in the BamHI site of pUC18 |
| anp02580 | Apr; Anabaena sp. chromosomal DNA from bp 4321373 to 4329600 in the BamHI site of pUC18 |
| anp03035 | Apr; Anabaena sp. chromosomal DNA from bp 4241824 to 4248722 in the BamHI site of pUC18 |
| anp03337 | Apr; Anabaena sp. chromosomal DNA from bp 1859242 to 1867412 in the BamHI site of pUC18 |
| anp04001 | Apr; Anabaena sp. chromosomal DNA from bp 4833192 to 4840653 in the BamHI site of pUC18 |
| anp04055 | Apr; Anabaena sp. chromosomal DNA from bp 1510438 to 1518550 in the BamHI site of pUC18 |
| anp04490 | Apr; Anabaena sp. chromosomal DNA from bp 1584150 to 1591582 in the BamHI site of pUC18 |
| pGEM-T Easy | Apr; cloning vector (Promega Corp.) |
| pK18 | Kmr; cloning vector (27) |
| pRL443 | Apr Tcr; conjugative plasmid (10) |
| pRL838 | Cmr Emr; BAC vector (18) (GenBank accession no. AF403425) |
| pRL2801 | Apr; internal fragment of all3850 amplified by PCR using genomic DNA from Anabaena sp. as template and primers IDT147 (5′-TAAATTCTCAGGCAGCAGATGA-3′) and IDT148 (5′-TGTAACACCGATGATTCTGGAG-3′), cloned in pGEM-T Easy |
| pRL2807 | Apr; internal fragment of alr1728 amplified by PCR using genomic DNA from Anabaena sp. as template and primers IDT161 (5′-AATCTCGCTACTCGCTTACCAG −3′) and IDT162 (5′-GTTTAGCATGACGGGGTTTAAG-3′), cloned in pGEM-T Easy |
| pRL2816a | Apr Cmr Emr; PstI fragment containing C.CE3-oriT cassette from pRL2665b (17) transferred to the PstI site of pRL2801 |
| pRL2821 | Apr Cmr Emr; PstI fragment containing C.CE3-oriT cassette from pRL2665b (17) transferred to the PstI site of pRL2807 |
| pRL2831a | Smr Spr; PglnA-containing RSF1010 derivative (17, 40) |
| pRL2831b | Smr Spr; same as pRL2831a, but PglnA oppositely oriented (17) |
| pRL2833a | Cmr Emr; PglnA-containing pDU1 derivative (10) |
| pRL2833b | Cmr Emr; same as pRL2833a, but PglnA oppositely oriented (10) |
| pRL2879 | Smr Spr; all1590, downstream of all1591, was excised as an EaeI-RsaI fragment of anp03337 and transferred between PspOMI and StuI of pRL2831a |
| pRL2888 | Smr Spr; all4018-containing XmnI-NheI fragment of anp04001 transferred between XbaI and StuI of pRL2831a |
| pRL3006 | Smr Spr; all3520-containing HpaI-Acc65I fragment of anp03035 transferred between StuI and BsiWI of pRL2831a |
| pRL3007 | Smr Spr; all1338-containing HincII-EaeI fragment of anp04490 transferred between StuI and PspOMI of pRL2831a |
| pRL3008 | Smr Spr; all3582-containing HindIII-HincII fragment of anp00676 transferred between HindIII and Ecl136II of pRL2831b |
| pRL3009 | Smr Spr; all4388-containing NlaIII-HincII fragment of anp02032 transferred between SphI and Ecl136II of pRL2831b |
| pRL3026 | Smr Spr; alr4311-containing Eco47III-NlaIII fragment of anp00427 transferred between Ecl136II and SphI of pRL2831b |
| pRL3038 | Smr Spr; all4019-containing SphI-HindIII fragment of anp04001 transferred between the same sites in pRL2831b |
| pRL3050 | Smr Spr; all3278-containing NheI fragment of anc1006 transferred into BlnI of pRL2831a |
| pRL3106 | Kmr; genes downstream of all3582 were excised as an XcmI (blunted)-PstI fragment of anp02580 and cloned between SmaI and PstI of pK18 |
| pRL3118 | Cmr Emr; genes downstream of all3582 were excised as a SacI-SphI fragment of pRL3106 and transferred between the same sites in pRL2833b |
| pRL3132 | Kmr; all4387-containing SpeI-Cac8I fragment of anp02032 cloned between XbaI and SmaI of pK18 |
| pRL3134 | Cmr Emr; genes downstream of all3520 were excised as a SapI (blunt)-BsrGI fragment of anc0756 and transferred between StuI and BsiWI of pRL2833a |
| pRL3142 | Cmr Emr; all4387-containing PstI-Acc65I fragment of pRL3132 transferred between NsiI and BsiWI of pRL2833a |
| pRL3155 | Apr Cmr Emr; genomic DNA of SR2821 was digested with HindIII, religated, and transformed to E. coli. Plasmids recovered on plates containing Cm contain pRL2821 and its neighboring genomic sequences |
| pRL3170 | Cmr Emr; all1591-containing EaeI-PvuII fragment of anp03337 transferred between PspOMI and StuI of pRL2833a |
| pRL3171 | Cmr Emr; all3850-containing PstI-DraI fragment of anp00920 transferred between NsiI and StuI of pRL2833a. This fragment extends 41 bp 3′ and 396 bp 5′ from all3850 (88 bp 3′ from the next upstream orf, asl3851) |
| pRL3172 | Cmr Emr; all4018-containing NsiI-XhoI fragment of pRL2888 transferred between the same sites of pRL2833a |
| pRL3180 | Smr Spr; asl3849-containing ApaI-XmnI fragment, which overlaps the 3′ ends of neighboring ORFs asr3848 and all3850, of anp00920 transferred between ApaI and StuI of pRL2831a |
| pRL3810 | Kmr Nmr; the coliphage T7 terminator as a BamHI-BglII fragment of pET-3 (28) inserted into the BamHI site at the end of the polylinker of pRL3040a (40) |
| pRL3811 | Kmr Nmr; SacI-XhoI fragment of pRL3171 transferred between the SacI and XhoI sites of pRL3810 |
| pUC18 | Apr; cloning vector (43) |
a
Drug resistance is indicated by a superscript r. The drugs are abbreviated as follows: Ap, ampicillin; Bm, bleomycin; Cm, chloramphenicol; Em, erythromycin; Km, kanamycin; Nm, neomycin; Sm, streptomycin; Sp, spectinomycin; Tc, tetracycline. Anabaena sp., Anabaena sp. strain PCC 7120; BAC, bacterial artificial chromosome.