Modification of the mycobacteriophage Ms6 attP core allows the integration of multiple vectors into different tRNAala T-loops in slow- and fast-growing mycobacteria

Table 1 Integration efficiency of vectors carrying attP core modifications

Vector	26 bp core *attP*^a	Integration efficiency/pAV6950^b		% integration into the tRNA^alaT-loop^c
		M. smegmatis	M. bovis BCG	M. smegmatis	M. bovis BCG
pAV6950	AGGGGTTCGAATCCCCTTAGCTCCAC	100%	100%	100% in tRNA^alaU	100% in tRNA^alaV
pBU	AGGGGTTCGAG TCCCCTTAGCTCCAC	1.1%	34%	ND	100% in alaU
pBT	AGGGGTTCGAG TCCCCTAG GCTCCAC	0.7%	18%	ND	100% in tRNA^alaT
pSV	AGGGGTTCGAT TCCCCTTAGCTCCAC	12%	0.8%	100% tRNA^alaV	ND
pST	AGGA GTTCGAATCT CCTAG GCTCCAC	0.05%	ND	ND	ND

a: Changes introduced by site-directed mutagenesis, with respect to the original attP site from pAV6950 are indicated in bold
b: Identical amounts of plasmid DNA were used to electroporate competent M. smegmatis or BCG. Integrants were numbered and the percentage of [integrants with modified attP/integrants with pAV6950] was calculated.
c: The insertion locus was determined by PCR amplification and the percentage of clones in the given tRNA^ala T-loop was calculated

ISSN: 1471-2199