Table 1.

Bacterial strains and plasmids used in this work

Strain or plasmidRelevant characteristic(s)Source or reference
A. vinelandii
 ATCC 9046Highly mucoidATCC
 AEIVMucoidSvein Valla
 WI12 algD::lacZ derivative of ATCC 9046, Kmr 5
 AC28 ampD::Tn5 derivative of WI12, Tcr This work
 ATD1ATCC 9046 with a nonpolar ampD::Gm mutationThis work
 ATE1ATCC 9046 with an ampE::Sp mutationThis work
 ATDE1ATCC 9046 with a ΔampDE::Gm mutationThis work
 AED1AEIV with a nonpolar ampD::Gm mutationThis work
 AEE1AEIV with anampE::Sp mutationThis work
 AEDE1AEIV with a ΔampDE::Gm mutationThis work
 AE28AEIV carrying anampD::Tn5 insertion, Tcr This work
 WID1WI12 with a nonpolarampD::Gm mutationThis work
 WIE1WI12 with an ampE::Sp mutationThis work
 WIDE1WI12 with a ΔampDE::Gm mutationThis work
E. coli
 DH5α supE44 ΔlacU169 hsdR17 recA1 endA1 gyrA96 thi-1 relA1 14
Plasmids
 pCP13RK2-derived cosmid vector; TcrKmr 7
 pBluescript SK+Plasmid used for subcloning DNA; Apr Stratagene
 pKT230Broad-host-range vector; SmrKmr 2
 pUT-mini-Tn5luxAB Suicide vector for mutagenesis with mini-Tn5luxAB; able to replicate only in strains expressing λ pir protein; Tcr 8
 pRK2013ColE1-tra (RK2); Kmr 11
 pHP45Ω-SpPlasmid used to provide an Ω Sprcassette 10
 pBSL141Plasmid used to provide a Gmr cassette 1
 pK28pBluescript SK+ with a 3.0-kb KpnI fragment containing the site of transposition in AC28; Tcr This work
 pSM194Cosmid containing 25 kb of A. vinelandii DNA including ampDE operon derived from pCP13; Tcr This work
 pS194.5pBluescript SK+ with a 2.0-kb SmaI fragment containing ampDEoperonThis work
 pS195pS194.5 derivative containingampD 5′ endThis work
 pS196pS194.5 derivative containing ampD gene and ampE 5′ endThis work
 pS196.2pS194.5 derivative containing ampE 5′ endThis work
 pSD1pS196 derivative containingampD::Gm mutationThis work
 pSE1pS196.2 derivative containingampE::Sp mutationThis work
 pSDE1pS194.5 derivative containing ΔampDE::Gm mutationThis work
 pKT194.5pKT230 derivative carrying ampDE operon; Smr This work