Identification of RanBMP interacting with Shigella flexneri IpaC invasin by two-hybrid system of yeast

Table 1 Strains and plasmids

	Relevant characteristics	Source
Strains
S. flexneri 2a, 2457T	Wild type, Nalr	Maurelli AT
E. coli DH5α	SupE44, ΔlacU169 (Φ80 lacZΔM15), hsdR17, recA1, endA1, gyrA96, thi-1, relA1, Nalr	Our lab
S. cerevisiae AH109	MATa, trp1-901, leu2-3, ura3-52, his3-200, gal4Δ, gal80Δ, LYS2::GAL1UAS-GAL1TATA-HIS3, GAL2UAS-GAL2TATA-ADE2, URA3::MEL1UAS-MEL1TATA-LacZ	Clontech
Plasmids
pGBKT7	GAL4 DNA-BD, TRP1, c-Myc epitope tag, Kmr Clontech
pGBKT-ipaC	GAL4 DNA-BD fusion of IpaC, TRP1, c-Myc epitope tag, Kmr	This study
pACT2	GAL4 DNA-AD, HA epitope, LEU2, Apr	Clontech
pLAM5'-1	GAL4 DNA-BD fusion of the human lamin C, TRP1, a false-positive detection plasmid, Apr	Clontech
p1	A positive library plasmid	This study

Table 2 Detection of transcriptional activity of bait fusion protein in AH109

Tranformed plasmids	Growth on SD His-	X-Gal
pGBKT-ipaC	-	white
pGBKT-ipaC + pACT2	-	white

Table 3 Verification of the interaction in positive clones

	Tranformed plasmids	β-Gal activity
1	p1	white
2	p1 + pGBKT7	white
3	p1 + pLAM5’-1	white
4	p1 + pGBKT-ipaC	blue