Effect of the potential triplex DNA region on the in vitro expression of bacterial beta-lactamase gene in superhelical recombinant plasmids.
The effect of a pyrimidine/purine-biased stretch which has the potential to form an unusual triplex DNA structure on gene expression has been analyzed by measuring the activity of beta-lactamase as a reporter gene in recombinant plasmids. The Escherichia coli transformant carrying the plasmid p7ERS which has a potential triplex DNA region expressed about twofold more beta-lactamase activity than that carrying the plasmid pUC19. Since the expression of beta-lactamase has been shown to be affected by template topology in vitro, this in vivo observation suggests that the inserted pyrimidine/purine-biased stretch modulates the topology of flanking regions by forming unusual DNA structure to keep the template at the superhelicity favorable for the expression of beta-lactamase.
Kato
M
M
Department of Life Science, College of Integrated Arts and Sciences, University of Osaka Prefecture.
Shimizu
N
N
D001483:Base Sequence; D002511:Cephalosporins / Q000378:metabolism; D004247:DNA / Q000235:genetics* / Q000648:ultrastructure; D004269:DNA, Bacterial / Q000235:genetics / Q000502:physiology / Q000648:ultrastructure*; D004274:DNA, Recombinant / Q000235:genetics*; D004278:DNA, Superhelical / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure; D066246:ErbB Receptors / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015870:Gene Expression / Q000235:genetics*; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D014176:Protein Biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics; D014169:Transformation, Bacterial / Q000235:genetics; D001618:beta-Lactamases / Q000235:genetics* / Q000378:metabolism
D002511:Cephalosporins; D004269:DNA, Bacterial; D004274:DNA, Recombinant; D004278:DNA, Superhelical; D012333:RNA, Messenger; C051802:triplex DNA; C034313:pyridine-2-azo-4-dimethylaniline cephalosporin; D004247:DNA; D066246:ErbB Receptors; D001618:beta-Lactamases