The instability of a recombinant plasmid, caused by a prokaryotic-like promoter within the eukaryotic insert, can be alleviated by expression of antisense RNA.
A region of the cauliflower mosaic virus genome was found to direct the expression of a nucleic-acid-binding protein in Escherichia coli. This protein is apparently toxic for the bacteria and leads to a destabilization of plasmids containing that region. Antisense RNA was used to diminish the unwanted expression and to stabilize the respective recombinant plasmids. The approach described may prove useful in other cases where problems with cloning of eukaryotic DNA arise.
Fütterer
J
J
Friedrich Miescher Institut, Basel, Switzerland.
Gordon
K
K
Pfeiffer
P
P
Hohn
T
T
D004262:DNA Restriction Enzymes; D004274:DNA, Recombinant / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D005814:Genes, Viral; D009029:Mosaic Viruses / Q000235:genetics*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012313:RNA / Q000235:genetics*; D016372:RNA, Antisense
D004274:DNA, Recombinant; D004268:DNA-Binding Proteins; D016372:RNA, Antisense; D012313:RNA; D004262:DNA Restriction Enzymes