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OverviewPhage-derived ΦC31 integrase represents an attractive tool for site-directed recombination in mammalian cells. Integration is based on recombination between attachment site attB and wt/pseudo-attP' site. Disadvantages are inefficient nonviral gene delivery and aberrant events (15% chromosomal rearrangements/deletions) within the host genome. The study's aim was to increase safety and efficiency of ΦC31 integrase. DNA binding domain was mutated by alanine scan, 22 mutants were evaluated for improved integration and intramolecular recombination. The combination of beneficial mutations in addition to optimization of the integrase plasmid dose enhanced integration efficiencies from 1.7 to 5.5-fold. Several mutants showed cell line-dependent integration activities. Excision assays between native attB/attP sites revealed 5 mutants with 2-fold enhanced activity. Enhanced recombination between attB and 3 described attP' sites (hot spots) in the mammalian genome assumed preferred specificity. 2 mutants showed similar integration activity as wt due to hFIX expression in mouse hepatocytes. Mutational analysis revealed an efficient approach for improvements of integration efficiency in vitro. Full Product DetailsAuthor: Raphael LiesnerPublisher: Sudwestdeutscher Verlag Fur Hochschulschriften AG Imprint: Sudwestdeutscher Verlag Fur Hochschulschriften AG Dimensions: Width: 15.20cm , Height: 0.90cm , Length: 22.90cm Weight: 0.222kg ISBN: 9783838123509ISBN 10: 3838123506 Pages: 144 Publication Date: 27 February 2011 Audience: General/trade , General Format: Paperback Publisher's Status: Active Availability: In stock We have confirmation that this item is in stock with the supplier. It will be ordered in for you and dispatched immediately. Table of ContentsReviewsAuthor InformationTab Content 6Author Website:Countries AvailableAll regions |