Generation of a synthetic mammalian promoter library by modification of sequences spacing transcription factor binding sites.
作者： Jens Tornøe , Philip Kusk , Teit E. Johansen , Peter R. Jensen
DOI: 10.1016/S0378-1119(02)00878-8
关键词:
摘要: The development of a set synthetic mammalian promoters with different specific activities is described. library based on promoter, JeT, constructed as 200 bp chimeric promoter built from fragments the viral SV40 early and human beta-actin ubiquitin C promoters. JeT was made by separating included consensus boxes same distances in base pairs found wild-type promoters, thus preserving transcription factor interaction. resulting shown to drive reporter expression high levels enhanced green fluorescent protein secreted alkaline phosphatase assays. By replacing sequences binding sites randomized length, sets new strengths, spanning 10-fold range transcriptional activity cell culture, obtained. measured each highly reproducible when tested HiB5 ARPE-19 culture.
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