Regulated expression of recombinant DNA
OwnerJaime Flores-Riveros, Jean Adamus +1inventors
InventorsJaime Flores-Riveros, Jean Adamus, Grazia Ambrosini
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Abstract
This invention relates to regulatable recombinant expression constructs that provide regulated gene expression of mammalian genes.
Claims31
A recombinant expression construct encoding a gene operably linked to a promoter comprising transcriptional control elements that are responsive to a small molecule regulator, wherein expression of said gene is regulated by contacting a cell containing the recombinant expression construct with the small molecule regulator, and wherein regulated expression is mediated by at least two DNA operator sequences inserted upstream of the start of translation and downstream from the promoter TATA site and the start of transcription.
A method for producing a recombinant promoter element under transcriptional control of a transcriptional regulator protein, comprising the step of digesting DNA encoding a promoter with a restriction enzyme having a recognition site proximal to an mRNA transcription initiation site, mixing the digested DNA with an excess of a double-stranded oligonucleotide encoding transcription regulatory sequence recognized by said transcriptional regulator protein, ligating the DNA and double-stranded oligonucleotide under conditions wherein at least two copies of the double-stranded oligonucleotide are incorporated at the restriction digestion site, and obtaining the recombinant promoter element.
Dependencies are inferred from the claim text.