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Stem Cell Related Patent Number US6221647
| Title: | Efficient construction of gene targeting using phage-plasmid recombination | | Inventors: | Woychik, Richard; Beechwood, OH, USA | | Summary: | This invention describes a system of production of gene targeting constructs in bacteria via homologous recombination between bacterial phage and plasmids. Such a method of construction involves the preparation of a probe plasmid comprising a marker cassette, which in turn comprises a suppressor t-RNA gene and a mammalian cell selectable marker. The marker cassette is flanked on at least one side by probe DNA, wherein the probe DNA comprises at least a portion of an exon of a gene to be targeted. Further disclosed are means of introducing the probe plasmid into the population of homologous, recombination-proficient, suppressor-free bacterial host cells. Also included are the steps for preparing a target phage, such that the target phage comprises at least one suppressible mutation in a gene necessary for phage growth and a target DNA consisting of a portion of a genomic region to be targeted, such that the target DNA is homologous to all or part of the probe DNA. Methods of infection, recombination and isolation are also provided. | | Abstract: | A method for producing gene targeting constructs in bacterial by way of homologous recombination between bacterial phage and plasmids. | | US Patent Website: | Click Here for Full Text of Patent | | Title Number: | US6221647 | | Application Number: | US1999000262445 | | Date Filed: | 04/03/1999 | | Date Published: | 24/04/2001 | | Assignee: | Amgen Inc., Thousand Oaks, CA, USA |
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