Expression, Purification, and Functional Analysis of Novel RelE Operon from X. nematophila

Table 2

Strains and plasmid used in this study.

Construct/strain	Characteristic	Source
E. coli DH5	supE44ΔlacU169 hsdR17 recA1 endA1 gyrA96 thi-1 relA1Φ80 dlacZ ΔM15	Invitrogen
E. coli BL21 (DE3) pLysS	F-ompT hsdSB(rB−mB−) gal dcm(DE3)plysS(CmR)	Novagen
E. coli TOP10	F-Φ80lacZΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17 (rK–, mK+) phoA supE44 -thi-1 gyrA96 relA1	Invitrogen
E. coli BL21 (DE3)	F-ompT hsdSB(rB–, mB–) gal dcm (DE3)	Novagen
pET 28 (a)	5.3 kb expression vector; kanr	Novagen
pBAD His (c)	4.1 kb, L-arabinose regulated pBR322-derived expression vectors designed for regulated, recombinant protein expression and purification in E. coli.	Invitrogen
pGEX4T1	4.96 kb, bacterial vector for expressing fusion proteins with a thrombin site, GST tagged	GE Healthcare
pJSL/JSL	pBAD His (c) alone without insert in E. coli Top 10 cells	Present study
pJSL1/JSL1	pGEX4T vector containing 285 bp RelB toxin gene from RelE TA module of X. nematophila.	Present study
pJSL2/JSL2	pET 28 (a) vector containing 252 bp RelE antitoxin gene from RelE TA module of X. nematophila.	Present study
pJSL3/JSL3	pBAD vector containing 285 bp RelE toxin gene from RelE TA module of X. nematophila.	Present study
pJSL4/JSL4	pBAD vector containing 526 bp full RelE operon from the genome of X. nematophila.	Present study