A new method for constructing linker scanning mutants

Episode von Medizin - Open Access LMU - Teil 06/22 von Ludwig-Maximilians-Universität München · 1. Januar 1987

A new procedure for the construction of linker scanning mutants is described. A plasmid containing the target DNA is randomly linearized and slightly shortened by a novel combination of established methods. After partial apurination with formic acid a specific nick or small gap is introduced at the apurinic site by exonuclease III, followed by nuclease S1 cleavage of the strand opposite the nick/gap. Synthetic linkers are ligated to the ends and plasmids having the linker inserted in the target DNA are enriched. Putative linker scanning mutants are identified by their topoisomer patterns after relaxation with topoisomerase I. This technique allows the distinction of plasmids differing in length by a single basepair. We have used this rapid and efficient strategy to generate a set of 32 linker scanning mutants covering the chicken lysozyme promoter from –208 to +15

Weitere Episoden von „Medizin - Open Access LMU - Teil 06/22“

• Adrenostatische Therapie mit Metyrapon und Aminoglutethimid beim ACTH-abhängigen Cushing-Syndrom 1. Januar 1987
• Ca2+-Stimulated Catecholamine Release from alpha-Toxin Permeabilized PC12 Cells 1. Januar 1987
• Changes in trabecular bone, hematopoiesis and bone marrow vessels in aplastic anemia, primary osteoporosis, and old age 1. Januar 1987
• Die Heidelberger Antiepileptikastudie 1. Januar 1987
• Dyskinesie der Abdominalmuskulatur bei Hypochondrie 1. Januar 1987
• Endocrine cells share expression of N-CAM with neurones 1. Januar 1987
• Estrone and estradiol concentrations in human ovaries, testes, and adrenals during the first two years of life 1. Januar 1987
• Further Characterization of Dopamine Release by Permeabilized PC 12 Cells 1. Januar 1987
• Grenzzoneninfarkte mit symptomatischer Epilepsie 1. Januar 1987
• Immunological characterization of chromogranins A and B and secretogranin II in the bovine pancreatic islet 1. Januar 1987
• Isolation and Characterization of cDNA Clones Encoding the Human Carcinoembryonic Antigen Reveal a Highly Conserved Repeating Structure 1. Januar 1987
• Mitochondrial Protein Import 1. Januar 1987
• Monoclonal antibodies against human astrocytomas and their reactivity pattern 1. Januar 1987
• Purification of alpha-toxin from Staphylococcus aureus and application to cell permeabilization 1. Januar 1987
• Tendenzen der Rechtsprechung zum Gebiet der Geburtshilfe 1. Januar 1987

Alle Episoden von Medizin - Open Access LMU - Teil 06/22 · Mehr Bildung-Podcasts