Tight control of gene expression in eucaryotic cells by tetracycline-responsive promoters
OwnerManfred Gossen, Hermann Bujardinventors
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Abstract
A first polynucleotide molecule coding for a transactivator fusion protein comprising the tet repressor and a protein capable of activating transcription in eucaryotes. A second polynucleotide molecule coding for a protein, wherein said polynucleotide is operably linked to a minimal promoter operably linked to at least one tet operator sequence is also disclosed. A method to regulate the expression of a protein coded for by a polynucleotide, by cultivating the eucaryotic cell of the invention in a medium comprising tetracycline or a tetracycline analogue is also disclosed. Kits containing the polynucleotide molecules are also disclosed.
Claims19
A polynucleotide molecule coding for a transactivator fusion protein comprising a prokaryotic tet repressor and a eucaryotic transcriptional activator protein domain.
A polynucleotide molecule coding for a protein, wherein said polynucleotide is operably linked to a minimal promoter and at least one tet operator sequence.
A eucaryotic cell transfected with
- (a) a first polynucleotide molecule coding for a transactivator fusion protein comprising a prokaryotic tet repressor and a eucaryotic transcriptional activator protein domain;
- and
- (b) a second polynucleotide molecule coding for a protein, wherein said second polynucleotide molecule is operably linked to a minimal promoter and at least one tet operator sequence.
A kit comprising a carrier means having in close confinement therein at least two container means, wherein a first container means contains a first polynucleotide molecule coding for a transactivator fusion protein comprising a prokaryotic tet repressor and a eucaryotic transcriptional activator protein domain, and a second container means contains a second polynucleotide molecule comprising a minimal promoter operably linked to at least one tet operator sequence, wherein said minimal promoter is capable of being ligated to a heterologous gene sequence coding for a polypeptide.
A kit comprising a carrier means having in close confinement therein at least two container means, wherein a first container means contains a eucaryotic cell transfected with a first polynucleotide molecule coding for a transactivator fusion protein comprising a prokaryotic tet repressor and a eucaryotic transcriptional activator protein domain, and a second container means contains a second polynucleotide molecule comprising a minimal promoter operably linked to at least one tet operator sequence, wherein said minimal promoter is capable of being ligated to a heterologous gene sequence coding for a polypeptide.
Dependencies are inferred from the claim text.