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- Publisher Website: 10.1101/pdb.prot5291
- Scopus: eid_2-s2.0-70949101008
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Article: Construction of gene-targeting vectors by recombineering
| Title | Construction of gene-targeting vectors by recombineering |
|---|---|
| Authors | |
| Issue Date | 2009 |
| Citation | Cold Spring Harbor Protocols, 2009, v. 4, n. 9 How to Cite? |
| Abstract | The critical step in generating a knockout mouse is gene-targeting vector construction. Recombineering technology has greatly streamlined the vector construction process. This protocol describes a method for making conditional knockout (cko) targeting vectors using the pSim18 plasmid. This plasmid carries the three λ phage Red genes (Gam, Bet, and Exo) under the control of the pL promoter, which is in turn regulated by the temperature-sensitive CI857 repressor. Hence, the heat-inducible recombineering functions can be easily delivered to bacterial artificial chromosomes (BACs) using a simple plasmid transformation, allowing one to manipulate any cloned mouse genomic region in Escherichia coli for cko vector construction. The conditional targeting vectors described in this protocol generate a flexible reporter/null/conditional allele in the mouse. Copyright © 2009 by Cold Spring Harbor Laboratory Press. |
| Persistent Identifier | http://hdl.handle.net/10722/249033 |
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.author | Lee, Song choon | - |
| dc.contributor.author | Liu, Pentao | - |
| dc.date.accessioned | 2017-10-27T05:58:55Z | - |
| dc.date.available | 2017-10-27T05:58:55Z | - |
| dc.date.issued | 2009 | - |
| dc.identifier.citation | Cold Spring Harbor Protocols, 2009, v. 4, n. 9 | - |
| dc.identifier.uri | http://hdl.handle.net/10722/249033 | - |
| dc.description.abstract | The critical step in generating a knockout mouse is gene-targeting vector construction. Recombineering technology has greatly streamlined the vector construction process. This protocol describes a method for making conditional knockout (cko) targeting vectors using the pSim18 plasmid. This plasmid carries the three λ phage Red genes (Gam, Bet, and Exo) under the control of the pL promoter, which is in turn regulated by the temperature-sensitive CI857 repressor. Hence, the heat-inducible recombineering functions can be easily delivered to bacterial artificial chromosomes (BACs) using a simple plasmid transformation, allowing one to manipulate any cloned mouse genomic region in Escherichia coli for cko vector construction. The conditional targeting vectors described in this protocol generate a flexible reporter/null/conditional allele in the mouse. Copyright © 2009 by Cold Spring Harbor Laboratory Press. | - |
| dc.language | eng | - |
| dc.relation.ispartof | Cold Spring Harbor Protocols | - |
| dc.title | Construction of gene-targeting vectors by recombineering | - |
| dc.type | Article | - |
| dc.description.nature | link_to_subscribed_fulltext | - |
| dc.identifier.doi | 10.1101/pdb.prot5291 | - |
| dc.identifier.scopus | eid_2-s2.0-70949101008 | - |
| dc.identifier.volume | 4 | - |
| dc.identifier.issue | 9 | - |
| dc.identifier.spage | null | - |
| dc.identifier.epage | null | - |
| dc.identifier.eissn | 1559-6095 | - |
| dc.identifier.issnl | 1559-6095 | - |