TABLE 2.

Bacterial strains, plasmids, and oligonucleotides

Designation	Description	Source or purpose
Strains and mutations
Rosetta 2 (DE3) pLysS	E. coli used for protein expression	Merck Millipore, Germany
DH5α	E. coli used for cloning	TaKaRa, Japan
T. thermophilus HB8	Wild type	BioResource Center, RIKEN, Japan.
ΔTTHA0328	TTHA0328 deletion strain of T. thermophilus	This work
Plasmids
pET11a-TTHA0328	Expression plasmid for nicotinamidase	BioResource Center, RIKEN, Japan
pUC19-HTK	Source of thermostable kanamycin resistance cassette	BioResource Center, RIKEN, Japan
pUC19-ΔTTHA0328	Plasmid for TTHA0328 deletion, containing HTK gene	This work
Oligonucleotides
U-F	5′-ATTCGAGCTCGGTACCCGGGGGAGACCCGGTCCCCGTA-3′	PCR for upstream region of TTHA0328
U-R	5′-GCCGTCAACGCACCTGGACCATGCCCTC-3′	PCR for upstream region of TTHA0328
D-F	5′-GGTCCAGGTGCGTTGACGGCGGATATGG-3′	PCR for downstream region of TTHA0328
D-R	5′-AAACAAACCGCGTAACCAACATGATTAAGAATTATTAGAGG-3′	PCR for downstream region of TTHA0328
HTK-F	5′-GTTGGTTACGCGGTTTGTTTGAGCACCCCG-3′	PCR for HTK
HTK-R	5′-CCTGCAGGTCGACTCTAGAGCCTCGCCGAAGGCGGGAA-3′	PCR for HTK
Seq_F	5′-GCGTTGTTGAAGAAGAGCTGGG-3′	PCR for sequencing
Seq_R	5′-CTTTAGGGGGAGGACAGGCTTC-3′	PCR for sequencing