Methods and materials for control of transcription by metals

C - Chemistry – Metallurgy – 12 – N

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195/1.2, 195/1.2

C12N 15/67 (2006.01) C12N 15/79 (2006.01)

Patent

CA 1318618

Abstract The transcription of DNA sequences in living cells is subjected to external regulation by incorporation of promoter/regulator DNA sequences responsive to metals. More particularly, regulation of the transcription of selected exogenous DNA sequences incorporated into eukaryotic host cells is facilitated by operative association (e.g., fusion) of the selected sequence to a promoter/regulator DNA sequence which is positively or negatively responsive to environmental variation in the concentration of heavy metal ions. As an example, a structural gene for thymidine kinase from herpes simplex virus, fused to the promoter/regulator DNA sequence of a mouse metallothionein-I gene and incorporated on a suitable DNA plasmid vector, is introduced into mouse embryos and stably incorporated therein. Gene expression in differentiated cells of adult mice resulting from the embryo is subsequently regulatable by administration of heavy metals such as cadmium. Metal regulatory elements (MREs) may be inserted into exogenous promoters in order to confer metal inducibility thereupon or to increase metal inducibility thereof. These MREs have the sequence TGCRCQCK wherein T is a thymidine nucleotide, G is a guanidine nucleotide, C is a cytidine nucleotide, R is a purine nucleotide, Q is a pyrimidine nucleotide or guanidine, and K is a purine nucleotide or cytidine.

523866

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