The Dominant Selective Marker APH 3′ and the Study of the Expression of the Cotransfected Gene
Molecular cloning and genetic engineering often lead to the assay of the biological properties and of the molecular integrity of purified clones of DNA. The reintroduction of cloned DNA material into a living eukaryotic cell should allow such a verification. Once the methodology for amplifying the DNA (usually through a bacterial plasmid) and for insuring the penetration of the DNA into the cell (transfection of a bacterial plasmid only or of a vectored eukaryotic recombinant) has been achieved, the discrimination of the cells that harbour or express the foreign gene must be brought about. A certain number of selective markers have been developed. The herpes simplex virus thymidine kinase (HSV TK) cloned in pBR322 (Colbère-Garapin et al., 1979; Bolivar et al., 1977) has been the first of such markers to be expressed in thymidine kinase defective (TK-) cells. An important aspect of such transfection has been the discovery, by Perucho et al. (1980) that when two genes are transfected together into a cell, they are integrated via a “pekalasome” at the same locus on a chromosome. This allows, with the help of a selective medium, the survival of cells which often coexpress the marker gene and the recombinant gene.
KeywordsThymidine Kinase Antigen Gene Thymidine Kinase Gene Bacterial Plasmid Dominant Selective Marker
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