TABLE 1.
Bacterial strains and plasmids used
| Strain or plasmid | Derivation and/or relevant characteristicsa |
|---|---|
| Strains of Anabaena sp. | |
| PCC 7120 | Wild type, from R. Haselkorn |
| DR1069 | Smr Spr; alr2835 (hepA) mutation resulting from double recombination with pRL1069 (5) |
| FQ57 | Bmr Nmr Smr; alr2827::Tn5-1063 |
| FQ344 | Bmr Nmr Smr; alr2841::Tn5-1063 |
| FQ428 | Bmr Nmr Smr; alr2837::Tn5-1063 |
| FQ470 | Bmr Nmr Smr; alr2833::Tn5-1063 |
| FQ610 | Bmr Nmr Smr; alr2838::Tn5-1063 |
| FQ773 | Bmr Nmr Smr; alr2832::Tn5-1063 |
| FQ794 | Bmr Nmr Smr; alr2839::Tn5-1063 |
| FQ885 | Bmr Nmr Smr; alr2825::Tn5-1063 |
| FQ1630 | Bmr Nmr Smr; alr2840::Tn5-1063 |
| FQ1631 | Bmr Nmr Smr; alr2831::Tn5-1063 |
| Strains of E. coli | |
| DH10B | 17 |
| DH5αMCR | Invitrogen Corp., received courtesy of J. C. Meeks |
| HB101 | 6 |
| Plasmids | |
| anc0293 | Cmr Emr; Anabaena sp. strain PCC 7120 chromosomal DNA from bp 3436903 to 3449816 in the BamHI site of pRL838 |
| anc0626 | Cmr Emr; Anabaena sp. strain PCC 7120 chromosomal DNA from bp 3451665 to 3468470 in the BamHI site of pRL838 |
| anc0901 | Cmr Emr; Anabaena sp. strain PCC 7120 chromosomal DNA from bp 3446424 to 3461510 in the BamHI site of pRL838 |
| anp02206 | Apr; Anabaena sp. strain PCC 7120 chromosomal DNA from bp 3440325 to 3447227 in the BamHI site of pUC18 |
| anp04627 | Apr; Anabaena sp. strain PCC 7120 chromosomal DNA from bp 3444424 to 3452207 in the BamHI site of pUC18 |
| anp08632 | Apr; Anabaena sp. strain PCC 7120 chromosomal DNA from bp 3457063 to 3463731 in the BamHI site of pUC18 |
| pGEM-T Easy | Apr; cloning vector (Promega Corp.) |
| pK18 | Kmr; cloning vector (28) |
| pRL443 | Apr Tcr; conjugative plasmid (13) |
| pRL838 | Cmr Emr; BAC vector (20) (GenBank accession no. AF403425) |
| pRL1124 | Kmr; methylase-encoding plasmid (13) |
| pRL1383a | Smr Spr; RSF1010-based cloning vector (GenBank accession no. AF403428) |
| pRL2665b | Apr Cmr Emr; source of C.CE3-oriT(RK2) cassette to make derivatives of pUC18 and pGEM-T Easy conjugable to, and selectable in, Anabaena sp. strain PCC 7120 (GenBank accession no. AY661563) |
| pRL2683a | Kmr Smr Spr; pRL1124 made mobilizable by the insertion, at its unique PstI site, of an oriT (RK2) fragment (32) in whose SalI site is inserted Smr Spr cassette C.S4 (3, 37) |
| pRL2686 | Kmr; Smr Spr marker removed from pRL2683a by SalI digestion and religation |
| pRL2792 | Apr; internal fragment of alr2833 amplified by PCR using genomic DNA from Anabaena sp. strain PCC 7120 as template and primers IDT145 and IDT146, cloned in pGEM-T Easy |
| pRL2815 | Apr Cmr Emr; PstI fragment containing C.CE3-oriT cassette from pRL2665b transferred to the PstI site of pRL2792 |
| pRL2831a | Smr Spr; glutamine synthetase promoter PglnA on a 0.7-kb EcoRI fragment from pRL559 (14) transferred to the EcoRI site of pRL1383a, oriented toward the BamHI site |
| pRL2831b | Smr Spr; same as pRL2831a, but PglnA oppositely oriented |
| pRL2862 | Smr Spr; alr2831-containing StuI-HindIII fragment of anc0293, subcloned between the Ecl136II and HindIII sites of pRL2831b |
| pRL2863 | Smr Spr; alr2832-containing EcoRV-XhoI fragment of anc0293 transferred between the StuI and XhoI sites of pRL2831a |
| pRL2864 | Smr Spr; alr2827-containing XmnI-XbaI fragment of anc0293, subcloned between the StuI and XbaI sites of pRL2831a |
| pRL2865a | Smr Spr; alr2833-containing HindIII fragment of anp04627 transferred to the HindIII site of pRL2831b |
| pRL2866 | Smr Spr; alr2841-containing HindIII fragment of anc0626, transferred to the HindIII site of pRL2831b |
| pRL2871 | Kmr; alr2825-containing KpnI-XmnI fragment of anp02206 transferred between the KpnI and HincII sites of pK18 |
| pRL2872 | Kmr; alr2839-containing EcoRV fragment of anp08632 inserted in the HincII site of pK18 |
| pRL2873 | Smr Spr; alr2840-containing HincII-XbaI fragment of anp08632, transferred between the StuI and XbaI sites of pRL2831a |
| pRL2875 | Smr Spr; alr2825-containing KpnI-SphI fragment of pRL2871, transferred between the same sites of pRL2831b |
| pRL2876 | Smr Spr; alr2839-containing KpnI-SphI fragment of pRL2872, transferred between the same sites of pRL2831b |
| pRL2877 | Smr Spr; alr2837-containing XbaI-BglII fragment of anp08632 transferred between the XbaI and BamHI sites of pRL2831a |
| pUC18 | Cloning vector (41) |
a
Sm, streptomycin; Bm, bleomycin; Cm, chloramphenicol; Ap, ampicillin; Tc, tetracycline; Km, kanamycin.