Identification of 3′-terminal exons from yeast artificial chromosomes

David B. Krizman, Tiffany A. Hofmann, Udaya DeSilva, Eric D. Green, Paul S. Meltzer, Jeffrey M. Trent

Research output: Contribution to journalArticle

Abstract

We report an extension of 3′-terminal exon trapping technology to the identification of transcribed sequences from yeast artificial chromosomes (YACs). A 350-kb YAC containing mouse genomic DNA was gel-purified and used as the target DNA for the 3′-terminal exon trapping strategy. A novel direct ligation/ transfection approach was employed to increase the efficiency of trapping 3′-terminal exons from recombinant vector-derived chimeric mRNA. The resulting RT-PCR product was then used to generate a plasmid library. Randomly chosen individual subclones from this library were sequenced, and the results indicate that 86% met sequence criteria characteristic of 3′-terminal exons, whereas 14% were background from identified sources. PCR mapping efforts suggest eight putative last exons present within this YAC, whereas RT-PCR studies demonstrate that three reside within valid expressed sequences.

Original languageEnglish (US)
Pages (from-to)322-326
Number of pages5
JournalGenome research
Volume4
Issue number6
DOIs
StatePublished - Jan 1 1995

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

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    Krizman, D. B., Hofmann, T. A., DeSilva, U., Green, E. D., Meltzer, P. S., & Trent, J. M. (1995). Identification of 3′-terminal exons from yeast artificial chromosomes. Genome research, 4(6), 322-326. https://doi.org/10.1101/gr.4.6.322