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A simple way of insertion of the transcription initiation signal in DNA vehicles
V. L. Drutsa, V. R. Kaberdin, O. N. Koroleva
A. N. Belozersky Laboratory of Molecular Biology and Bioorganic Chemistry and Chemical Department of Moscow State University, Moscow 119899
Abstract: A new effective method of site-specific mutagenesis in the close vicinity of unique restriction sites of plasmids, based on the use of two oligonucleotide primers, mutagenic and auxiliary, has been suggested. A site-specific insertion of Pribnow box (TATAATG) before promoterless gal operon of the promoter-testing plasmid pHD-001-14-11 has been performed with the yield of mutants up to 95%. Data on the gal operon expression and S1-nuclease mapping of the transcription start point indicate the formation of an active promoter in the region of the insertion.
Russian Journal of Bioorganic Chemistry 1991, 17 (7):945-952