[A model prokaryotic promotor. III. Cloning and study of functional activity in vivo].
The efficiency of two schemes of oligonucleotide-directed insertion mutagenesis was studied in comparison with standard cloning of DNA duplexes based on blunt-end ligation. Using new approaches, the 30-bp consensus-like prokaryotic promoter was inserted in proper orientation in a promoter-testing plasmid at an almost 100% frequency. Data on the marker gal operon expression and S1-nuclease mapping of the transcription initiation points indicate formation of an active promoter in the region of the insertion.
Koroleva
O N
ON
Shilov
I A
IA
Sergeev
V N
VN
Drutsa
V L
VL
D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010957:Plasmids; D011387:Prokaryotic Cells; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
D004247:DNA
Model'nyĭ prokarioticheskiĭ promotor. III. Klonirovanie i issledovanie funktsional'noĭ aktivnosti in vivo.