[en] We describe a modification of the pAR3040 vector which results in its efficient stabilization during cell division. The parB locus of the plasmid R1 was introduced into the plasmid, pAR3040, to construct the pARHS vectors. These vectors are stable for at least 60 cell generations, even in the absence of selection by an antibiotic present in the culture media, both with or without IPTG induction.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
De Moerlooze, L.
Struman, Ingrid ; Université de Liège - ULiège > Département des sciences de la vie > Biologie et génétique moléculaire
Renard, Andre
Martial, Joseph ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire
Language :
English
Title :
Stabilization of T7-Promoter-Based Parhs Expression Vectors Using the Parb Locus
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Bibliography
Gerdes (1988) The parB (hok/sok) locus of plasmid R1: a general purpose plasmid stabilization system. Biotechnology 6:1402-1405.
Rosenberg, Lade, Chui, Lin, Dunn, Studier (1987) Vectors for selective expression of cloned DNAs by T7 RNA polymerase. Gene 56:125-135.
Sambrook, Fritsch, Maniatis Molecular Cloning. A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; 1989.
Studier, Moffatt (1986) Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J. Mol. Biol. 189:113-130.
Studier, Rosenberg, Dunn, Dubendorff (1990) Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 185:60-89.
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