Table 1 Bacterial strains, plasmids and oligonucleotides used in this study

E. coli OmniMAX

F′ {proAB lacI^q lacZΔM15 Tn10(Tet^R) Δ(ccdAB)} mcrA Δ(mrr hsdRMS-mcrBC) Φ 80(lacZ)ΔM15 Δ(lacZYA-argF)U169 endA1 recA1 supE44 thi-1 gyrA96 relA1 tonA panD; used for cloning

Invitrogen

B. subtilis 1012

leuA8 metB5 trpC2 hsrM1

[18]

pHCMC02-bgaB

PlepA-bgaB, inducer-free expression plasmid

[12]

pNDH33-bgaB

Pgrac01-bgaB, inducible

[3]

pLacI

Expression of LacI, used with E. coli cloning strain

Novagen

pHT01

Pgrac01, inducible

[9]

pHT01-bgaB

Pgrac01-bgaB, inducible

[9]

pHT24

Pgrac01, Strep-tag II, inducible

[9]

pHT10-gfp+

Pgrac01-gfp+, inducible

[9]

pHT100

Pgrac100-bgaB, inducible

[4]

pHT255

Pgrac100, inducible

[4]

pHT1168

Pgrac100-gfp+, inducible

[4]

pHT1650

Pgrac01-gfp+, ΔlacI, lacO1-lacO3 548 bp

This study

pHT1651

Pgrac01-gfp+, ΔlacI, lacO1-lacO3 787 bp

This study

pHT1653

Pgrac01-bgaB, ΔlacI, lacO1-lacO3 787 bp

This study

pHT1654

Pgrac100-bgaB, ΔlacI, lacO1-lacO3 732 bp

This study

pHT1655

Pgrac01-bgaB, ΔlacI, lacO1-lacO3 143 bp

This study

pHT1656

Pgrac100-bgaB, ΔlacI, lacO1-lacO3 88 bp

This study

pHT1657

Basic plasmid, used to construct pHT1655

This study

pHT1658

Basic plasmid, used to construct pHT1656, pHT1681, pHT1682

This stuy

pHT1660

Pgrac01-bgaB, ΔlacI, lacO1-lacO3 548 bp

This study

pHT1661

Basic plasmid, used to construct pHT1660

This study

pHT1674

Pgrac100-bgaB, ΔlacI, lacO1-lacO3 493 bp

This study

pHT1695

Pgrac100-gfp+, ΔlacI, lacO1-lacO3 493 bp

This study

pHT1696

Pgrac100-gfp+, ΔlacI, lacO1-lacO3 732 bp

This study

pHT2071

Pgrac01-bgaB, ΔlacI, lacO1-lacO3 280 bp

This study

pHT2079

Pgrac100-bgaB, ΔlacI, lacO1-lacO3 255 bp

This study

ON224

CCGGATGACGTCGAATTCTAAACCTTCCCGGCTTCATCATG

pHT1655, pHT1660, pHT1656

ON925

GAATTAGCTTGGTACCAAAGGAGGTAAGGATCACTAG

To amplify promoter Pgrac100 to construct pHT1674

ON926B

GACGTCGACTCTAGACATGGATCCTTCCTCCTTTAATTGG

To amplify promoter Pgrac100 to construct pHT1674

ON941

AAAGGAGGAAGGATCCATGAATGTGTTATC

pHT1655, pHT1660, pHT1656

ON985

CAATTGCGTTGCGCTCACTGCCGGTAC

pHT1657, pHT1658

ON986

CGGCAGTGAGCGCAACGCAATTGAGCT

pHT1657

ON1975

CAATTGCGTTGCGCTCACTGCCAGCGCT

To create lacO3 in pHT2071

ON1976

AGCGCTGGCAGTGAGCGCAACGCAATTGAGCT