TABLE 2.

Oligonucleotides designed for PCR amplification of the NHase α- and β-subunit genes and the amidase gene and for sequencing the NHase gene^a

Primer	Sequence	Direction of reading	Positionb	Usec
NHase primers
αNH1	5′-GTG AAC CAG ATG TCA GTA ACG ATC G	Forward	1907	Amp
αNH2	5′-CGC TCA GGC AGT CCT TGG TGA CG	Reverse	2507	Amp
βNH1	5′-GCA CAC CAT GGA TGG AGT ACA CG	Forward	2559	Amp
βNH2	5′-GCA GGC TCG AGG TAA CCC TCG	Reverse	3197	Amp
αNH4	5′-TCT ACG ACA CCA CCG CCG AAA CT	Forward	2394	Seq
βNH4	5′-CGA TGA CGT ACC TCT CGT AGT ACG	Reverse	2800	Seq
SV1	5′-GAC CAT GAT TTC CAG TGT CC	Reverse	3308	Seq
SV2	5′-GGC GAA GAA CCG ACG TAC C	Forward	3088	Seq
SV3	5′-GCC CGC ATA AGA AAA GGT GAA	Forward	1891	Amp
SV4	5′-TCA AAC GGT CTG GTC GGT ATA	Reverse	1328	Amp
Amidase primers
amd1	5′-GTG AAG CCG ATC ACA TCA GGA GC	Forward	245	Amp
amd2	5′-CGG GTA CCA ATC CCT TAC CGT CG	Reverse	2027	Amp

^aSee Materials and Methods for details.

^bPosition in the sequences of the Rhodococcus sp. strain N-774 NHase and amidase genes (30) except for the position of SV4, which is the position in the sequence of the Rhodococcus sp. strain N-774 NHase β-subunit downstream region Orf1188 (29). The DDBJ/EMBL/GenBank accession number for the NHase and amidase genes is X54074 and the accession number for Orf1188 is D30033.

^cAmp, amplification; Seq, sequencing.