Construction of Gene-Targeting Vectors by Recombineering
نویسندگان
چکیده
منابع مشابه
Construction of gene-targeting vectors by recombineering.
Recombineering is a technology that utilizes the efficient homologous recombination functions encoded by gamma phage to manipulate DNA in Escherichia coli. Construction of knockout vectors has been greatly facilitated by recombineering as it allows one to choose any genomic region to manipulate. We describe here an efficient recombineering-based protocol for making mouse conditional knockout ta...
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Gene targeting refers to the precise modification of a genetic locus using homologous recombination. The generation of novel cell lines and transgenic mouse models using this method necessitates the construction of a 'targeting' vector, which contains homologous DNA sequences to the target gene, and has for many years been a limiting step in the process. Vector construction can be performed in ...
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We describe a highly redundant murine genomic library in a new λ phage, λ knockout shuttle (λKOS) that facilitates the very rapid construction of replacement-type gene targeting vectors. The library consists of 94 individually amplified subpools, each containing an average of 40 000 independent genomic clones. The subpools are arrayed into a 96-well format that allows a PCR-based efficient reco...
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A rate-determining step in gene targeting is the generation of the targeting vector. We have developed bacteriophage gene targeting vectorology, which shortens the timeline of targeting vector construction. Using retro-recombination screening, we can rapidly isolate targeting vectors from an embryonic stem cell genomic library via integrative and excisive recombination. We have demonstrated tha...
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Objective(s): Up to now, many researches have been performed to improve the antitumoral effect of melanoma differentiation-associated gene-7 (mda-7) protein. The purpose of our research was to construct 3 expression vectors producing mda-7 in fusion with RGD (Arginine-Glycine-Aspartic acid) peptide and evaluate their expression. Materials and Methods: mda-7 gene with two different RGD sequ...
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ژورنال
عنوان ژورنال: Cold Spring Harbor Protocols
سال: 2009
ISSN: 1559-6095
DOI: 10.1101/pdb.prot5291