Generation of Random Internal Deletion Derivatives of YACs by Homologous Targeting to Alu Sequences

Jaemog Soh, Thomas M. Mariano, Gary Bradshaw, Robert J. Donnelly, Sidney Pestka

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

To facilitate the manipulation of human genomic DNA in yeast artificial chromosome (YAC) clones, a plasmid to integrate the selective marker for antibiotic G418 resistance into YACs and to delete some of the human DNA fragments from YACs was constructed. The linearized integration/deletion plasmid, which contains Alu family sequences at both ends, can recombine with YACs containing human repetitive sequences via homologous recombination. The homologous recombination results in a random integration of the antibiotic G418-resistant gene into a human genomic Alu sequence, and in most cases, an internal deletion within the YAC. The YACs with internal deletions can be useful to identify the location of the genes if they produce functional knockouts. In those cases when the integration/deletion event disrupts the integrity of the gene so it no longer can produce a viable and functional mRNA in fused eukaryotic cells, the site of integration in the YAC thus serves as a marker for the inactivated gene. In this report we describe a model system to locate specific genes in YACs.

Original languageEnglish (US)
Pages (from-to)301-309
Number of pages9
JournalDNA and Cell Biology
Volume13
Issue number3
DOIs
StatePublished - Mar 1994

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics
  • Cell Biology

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