Recombinant protein production in high cell density cultures of Escherichia coli with galactose as a gratuitous inducer.
A new expression system was developed by introducing two major modifications into the genome of Escherichia coli: a deletion in the gal operon (DeltagalEKT) to allow the use of the inexpensive compound galactose as a gratuitous inducer and the introduction of the gal P2 promoter driving the expression of the T7 RNA polymerase. The novel JRR10 strain containing these two features gives high-level expression of a reporter gene cloned under the T7 phi10 promoter in high cell density cultures. The cost of the induction of this novel system is more than 30 times lower than that of the IPTG-induced system of the widely used BL21 strain.
Microbiology Division, IBR (Instituto de Biología Molecular y Celular de Rosario), Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad Nacional de Rosario, Suipacha 531, Rosario, Argentina.
Hugo G
HG
Menzella
Hugo C
HC
Gramajo
D001483:Base Sequence; D017931:DNA Primers; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005690:Galactose / Q000378:metabolism*; D016680:Genome, Bacterial; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D014764:Viral Proteins
D017931:DNA Primers; D011994:Recombinant Proteins; D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases; D005690:Galactose