CLONING-FREE METHOD OF GENERATING TRANSCRIPTIONALLY AND POST-TRANSCRIPTIONALLY CONTROLLABLE EXPRESSION ACTIVE LINEAR REPORTER CONSTRUCTS

摘要

The present invention relates to regulated gene expression and reporter gene technology. It discloses a simple cloning-free method of generating a transcriptionally and/or post-transcriptionally controllable expression active linear reporter or gene construct with any desired introduced sequences. This method comprises a PCR amplification of a source DNA comprising a reporter or gene DNA sequence using (i) a forward primer comprising a first sequence part at the 3' end complementary to a region of the source DNA upstream of the reporter or gene DNA sequence, and a second sequence part at the 5' end comprising one or more introduced sequence(s), preferably transcriptional control sequence(s); and (ii) a reverse primer complementary to a region of the source DNA downstream of the reporter or gene DNA sequence.