. 2004 Oct;186(20):6944–6955. doi: 10.1128/JB.186.20.6944-6955.2004

TABLE 3.

Plasmids constructed in this study^a

Plasmid	Vector	Marker	Promoter	Gene	Expression^b	Fluorescence^c	Localization^d
pTK150	pEYFP	Ap^r	p65	p65-eyfp	NT	NT	NT
pTK153	pEYFP	Ap^r	lac (E. coli)	eyfp-p65	NT	NT	NT
pTK155	pISM2062.2	Ap^r Gm^r	p65	p65-eyfp	+	+	+
pTK158	pISM2062.2	Ap^r Gm^r	lac (E. coli)	eyfp-p65	−	−	−
pTK161	pISM2062.2	Ap^r Gm^r	p65	eyfp-p65	+	+	+
pTK162	pISM2062.2	Ap^r Gm^r	tuf	eyfp-p65	++	++	+
pTK164	pISM2062.2	Ap^r Gm^r	p65	eyfp	+	+	−
pTK164-D	pISM2062.2	Ap^r Gm^r Cm^r	p65	eyfp	NT	NT	NT
pTK165	pISM2062.2	Ap^r Gm^r	tuf	eyfp	++	++	−
pTK165-D	pISM2062.2	Ap^r Gm^r Cm^r	tuf	eyfp	NT	NT	NT
pTK205	pKV104	Ap^r Cm^r	lac (E. coli)	ecfp	NT	NT	NT
pTK207	pKV104	Ap^r Cm^r	tuf	ecfp	++	++	−
pTK207-D	pKV104	Ap^r Cm^r	tuf	ecfp	NT	NT	NT
pTK210	pTK207-D	Ap^r Cm^r	tuf	ecfp-p65	++	++	+
pMPN310-E	pENTR/D-TOPO	Km^r	None	hmw2	NT	NT	NT
pMPN311-E	pDONR201	Km^r	None	p41	NT	NT	NT
pMPN312-E	pDONR201	Km^r	None	p24	NT	NT	NT
pMPN310	pTK164-D	Ap^r Gm^r	p65	eyfp-hmw2	+	+	+
pMPN311	pTK164-D	Ap^r Gm^r	p65	eyfp-p41	+	+	+
pMPN312	pTK164-D	Ap^r Gm^r	p65	eyfp-p24	+	+	+
pMPN310-tuf	pTK165-D	Ap^r Gm^r	tuf	eyfp-hmw2	++	++	+
pMPN311-tuf	pTK165-D	Ap^r Gm^r	tuf	eyfp-p41	++	++	+
pMPN312-tuf	pTK165-D	Ap^r Gm^r	tuf	eyfp-p24	++	++	+

M. pneumoniae strains transformed with Tn4001 plasmids were analyzed by Western blotting and fluorescence microscopy. NT, not applicable or not tested.

Expression of the fusion protein in M. pneumoniae cells was monitored by Western blotting: +, ++, and −, moderate, strong, and no expression, respectively.

Fluorescence intensity was observed by microscopy: +, ++, and −, faint, strong, and no fluorescence, respectively.

Localization of the fusion protein at cell poles was observed by microscopy; +, present; −, absent.