Table 2.

Strains, plasmids, and primers used in this study

Bacterial strain, plasmid or primer	Relevant characteristic or sequence	Source or restriction
Strains
B. subtilis ACR	B. subtilis 168 with pMA5-HpaII-acr, source of yodC and ldhA genes	Lab stock
B. subtilis AY	B. subtilis ACR blocked in YodC by insertion of a Cat expression cassette	This study
B. subtilis AFY	B. subtilis ACR blocked in YodC by insertion of fdh gene	This study
B. subtilis AFYL	B. subtilis AFY blocked in LdhA by insertion of a CAT expression cassette	This study
E. coli JM109	recA1, endA1, gyrA96, thi-1, hsd R17(rk- mk +)supE44	Invitrogen
Candida boidinii	Source of fdh gene	Lab stock
Plasmids
pMD18-T	Cloning vector	Takara
pBGSC6	Containing Cat expression cassette	Lab stock
pMA5-HpaII	Expression vector (in E. coli, Apr; in B. subtilis, Kanr)	Lab stock
T-cat	pMD18-T with Cat expression cassette	This study
T-yodC	pMD18-T with yodC	This study
T-yodC::cat	cat inserted into the EcoR V site of T-yodC	This study
T-fdh	pMD18-T with fdh	This study
pMA5-fdh	pMA5-HpaII with fdh
T-HpaII-fdh	pMD18-T with HpaII-fdh	This study
T-yodC::fdh	fdh inserted into the Eco47 III site of T-yodC	This study
T-ldhA	pMD18-T with ldhA	This study
T-ldhA::cat	cat inserted into the EcoR V site of T-ldhA	This study
Primers 5′-3′
P1	ACCGGGATCCATGACGAATACTCTGGAT	BamH I
P2	ACCGACGCGTTTACAGCCAAGTTGATAC	Mlu I
P3	CGCGATATCAAAAAAGGATTGATTCTAATG	EcoR V
P4	CGCGATATCTAGTGACATTAGAAAACCGAC	EcoR V
P5	CGCCATATGATGAAGATCGTTTTAGTC	Nde I
P6	CGCACGCGTTTATTTCTTATCGTGTTTAC	Mlu I
P7	ACCGAGCGCTATTTTTTGAGTGATCTTCTC	Eco47 III
P8	ACCGAGCGCTTTATTTCTTATCGTGTTTAC	Eco47 III
P9	ACCGGGATCCATGATGAACAAACATG	BamH I
P10	ACCGACGCGTTTAGTTGACTTTTTGTTCTG	Mlu I

Underlined nucleotides are the restriction enzyme sites