National Research Council of Canada

Alaka Mullick

Bernard Massie

Peter C K Lau

Yasuo Konishi

Félix Malenfant

Carlos B Miguez

Young-Jun Choi

Yan Xu

Peter Ck Lau

Recently, the development of inducible expression systems has involved exploitation of the p-cym operon from. Disclosed herein are novel expression systems and components thereof, which involve the development of CymR variants with reverse DNA binding activity, such that they exhibit increased affinity for DNA in a presence rather than an absence of an effector molecule such as cumene or an equivalent thereof. Also disclosed are the CymR variants, fusion proteins incorporating such variants, and their use in the control and expression of polynucleotides.

Reverse cumate repressor mutant

Recently, the development of inducible expression systems has involved exploitation of the p-cym operon from. Disclosed herein are novel expression systems and components thereof, which involve the development of a CymR variant with reverse DNA binding activity, such that they exhibit increased affinity for DNA in a presence rather than an absence of an effector molecule such as cumene or an equivalent thereof. Also disclosed are the CymR variant, fusion proteins incorporating such a variant, and its use in the control and expression of polynucleotides. The CymR variant comprises a 142Glu to 142Gly single point mutation of wild type CymR.

Methylotrophic or methanotrophic bacteria such asare transformed with a gene of interest, and expression of the gene is regulated by means of a cumate repressor protein and an operator sequence which is operatively linked to the gene of interest, and the addition of an external agent. Specifically, the cymR repressor and cmt operator frommay serve to regulate gene expression in methylotrophic or methanotrophic bacteria with the addition of cumate.

Regulation of heterologous recombinant protein expression in methylotrophic and methanotrophic bacteria

The invention relates to a new 'gene-switch' (cumate-inducible switch) for mammalian cells, having a mammalian promoter which has a TATA element and is linked to the coding sequence of CymR. This switch is as useful in the development of expression systems and cell-based assays for functional genomics as in the generation of viral vectors for gene therapy.

DNA for eukaryotic expression of CymR

The invention relates to a new 'gene-switch' (cumate-inducible switch) for mammalian cells. This switch is as useful in the development of expression systems and cell-based assays for functional genomics as in the generation of viral vectors for gene therapy.

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Montreal, Canada

Alaka Mullick