Rescue of transfected genes from mammalian cells by functional selection in Escherichia coli |
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Authors: | Michael Strauss Udo Klessling and Ruth Kachler |
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Institution: | (1) Zentralinstitut für Molekularbiologie, Abteilung Virologie, Akademie der Wissenschaften der DDR, Robert-Rössle-Straße 10, DDR-1115 Berlin-Buch, German Democratic Republic |
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Abstract: | Summary We have established procedures for reisolating a transfected gene from mammalian cells by selection in Escherichia coli for the function of the gene product using the Herpes simplex virus thymidine kinase gene as a model. Rescue of the gene is accomplished by three different methods. The tk gene is recloned into a plasmid in which it is hooked up by either the lac promoter or a lac/tk hybrid promoter, or the original plasmid is cut out of the host cell DNA.As the lac/tk hybrid gene can be expressed and selected both in the mammalian and E. coli cells, this type of gene rescue allows investigations on mutagenesis and methylation processes. Additionally, it offers a simple way of studying the integration of the transfected gene into the mammalian genome.Abbreviations Ap
ampicillin
- FU
fluorouracil
- HSV
Herpes simplex virus |
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