Methods comprising apoptosis inhibitors for the generation of transgenic pigs

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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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