pPAC-ResQ: A yeast-bacterial shuttle vector for capturing inserts from P1 and PAC clones by recombinogenic targeted cloning

Jaya Bhargava, Cooduvalli S. Shashikant, Janet L. Carr, Kevin L. Bentley, Chris T. Amemiya, Frank H. Ruddle

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

We have developed a method to capture inserts from P1 and P1 artificial chromosome (PAC) clones into a yeast-bacteria shuttle vector by using recombinogenic targeting. We have engineered a vector, pPAC-ResQ, a derivative of pClasper, which was previously used to capture inserts from yeast artificial chromosome clones, pPAC-ResQ contains DNA fragments flanking the inserts in P1 and PAC vectors as recombinogenic ends. When linearized pPAC-ResQ vector and P1 or PAC DNA are cotransformed into yeast, recombination between the two leads to the transfer of inserts into pPAC- ResQ. pPAC-ResQ clones thus obtained can be further modified in yeast for functional analysis and shuttled to Escherichia coli to produce large quantities of cloned DNA. This approach provides a rapid method to modify P1/PAC clones for functional analysis.

Original languageEnglish (US)
Pages (from-to)337-339
Number of pages3
JournalGenomics
Volume56
Issue number3
DOIs
StatePublished - Mar 15 1999

All Science Journal Classification (ASJC) codes

  • Genetics

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