Characterization of a gene regulating antibiotic production in Bacillus subtilis BSD-2

Figures & data

Table 1. Strains and plasmids used in this study.

Strain or plasmid	Characteristics^a	Source
Strain
B. subtilis BSD-2 (CGMCC No.8440)		This lab
E. coli DH5α (ATCC 25 922)		This lab
Plasmid
pHV1249	Amp^r, Cm^r, Em^r	Guo Qinggang's lab
pBIUSKM	Amp^r	This lab
pEASY –T1	Amp^r, kana^r	This lab

^a Amp^r, ampicillin resistance: Cm^r, chloramphenicol resistance; Em^r, erythromycin resistance; kana^r, kanamycin resistance.

Table 2. Primers for PCR.

Primers	Sequence	Function or restriction sites
Tn10-5′	5′-CTTAGTGACAAGGGTGAT-3′	mini-Tn10 transposon sequence 5′ end primer
Tn10-3′	5′-TGCTGTAATAATGGGTAG-3′	mini-Tn10 transposon sequence 3′ end primer
SP1	5′-AACGTATTGCCCAGATCCGCCTA-3′	Chromosome-walking outside primer
SP2	5′-GAAACAAGGTTGGGACAAGCAC-3′	Chromosome-walking middle primer
SP3	5′-TTTTGCGGCATTCTGGCTACAC-3′	Chromosome-walking internal primer
P1	5′-ACGGGCCCATGCAAGGGAATCTGACTGCAC-3′	ApaI
P2	5′-GCGAGCTCTTATGCGCCCTTTTGATTT-3′	SacI
P3	5′-CGAAGCTTATGATTGAACAAGATGGATTGC-3′	HindIII
P4	5′-CGGGATCCATTCAGAAGAACTCGTCAAG-3′	BamHI

Figure 1. Bacillus subtilis mutants challenged with flat grey mould.

Figure 2 . Cloned mini-Tn10 transposon insertion site flanking sequences.

Table 3. Gene function of identified mutants.

Strains	Gene	Function
1–18, 1–9	ybf B	Acid transporter protein
1–24	between ybcL and ybcM	Does not encode proteins
1–19, 2–59	yod F	Na^+/metabolite permease

Figure 3. Effects of exposing recombinant strains to Botrytis cinerea; 1–2, recombinant strains.

Figure 4. Effects of recovery strains on grey mould; 1–4, recovery strains.