amiA is a negative regulator of acetamidase expression in Mycobacterium smegmatis

BMC Microbiology

Table 1 Promoter assay constructs.

pAGAN	pJEM	Primer	Primer	PCR product
number	number	name	sequence	position
120/121	12	PACT4	GGATCCATCTACCGGACTGCC	1677–1986
		PACT5	GGATCCGAAGGGACCGCAGTG
130/131	14/13	PACT6	GGATCCACCAGCCCGACG	1291–1536
		PACT7	GGATCCTGAGCGACGAGT
160/161	13	PACT10	GGATCCCCGAGCGCGATG	2000–2254
		PACT13	GGATCCACGTAGAAGAGC
170/171	12	PACT9	GGATCCCTCCTTGCACTC	2758–3026
		PACT12	GGATCCCGCCGTGATCGG

The regions shown in figure 3 were PCR-amplified from genomic DNA and cloned into pGEM-T (Promega). Primers were designed to contain Bam HI sites at each end. The fragments were then excised as Bam HI fragments and cloned into one of the pJEM vectors to give in-frame fusions to the promoterless β-galactosidase gene. The position of the PCR product with respect to the complete acetamidase region is given (assembled from Accession numbers U63095 and X57175).

ISSN: 1471-2180