EcoRI XhoIPstI EcoRI

UR DR

EcoRI XhoI

PstI EcoRI EcoRI XhoI

PstI EcoRI

UR DR

DRUR

pJQ200SK

PstI XhoI

Ori V (p15A)

Mob (RP4)

sacB

Gm

EcoRI EcoRI

pCRS530

GNm

GNm

PstI XhoI

EcoRI EcoRIGNm

Cut by EcoRI

Cut by PstI + XhoI

+ +

+

Quadruple Ligation

Strategy to construct gene specific inactivation vector

Gene specific inactivation vector

gusA nptII

See references for pJQ200SK and pCRS530 at the end of this presentation