Identification of 3'-terminal exons from yeast artificial chromosomes

D. B. Krizman, T. A. Hofmann, U. DeSilva, E. D. Green, P. S. Meltzer, J. 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
StatePublished - 1995
Externally publishedYes

Fingerprint

Yeast Artificial Chromosomes
Exons
Polymerase Chain Reaction
DNA
Libraries
Transfection
Ligation
Plasmids
Gels
Technology
Messenger RNA

ASJC Scopus subject areas

  • Molecular Biology

Cite this

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.

Identification of 3'-terminal exons from yeast artificial chromosomes. / Krizman, D. B.; Hofmann, T. A.; DeSilva, U.; Green, E. D.; Meltzer, P. S.; Trent, J. M.

In: Genome Research, Vol. 4, No. 6, 1995, p. 322-326.

Research output: Contribution to journalArticle

Krizman, DB, Hofmann, TA, DeSilva, U, Green, ED, Meltzer, PS & Trent, JM 1995, 'Identification of 3'-terminal exons from yeast artificial chromosomes', Genome Research, vol. 4, no. 6, pp. 322-326.
Krizman DB, Hofmann TA, DeSilva U, Green ED, Meltzer PS, Trent JM. Identification of 3'-terminal exons from yeast artificial chromosomes. Genome Research. 1995;4(6):322-326.
Krizman, D. B. ; Hofmann, T. A. ; DeSilva, U. ; Green, E. D. ; Meltzer, P. S. ; Trent, J. M. / Identification of 3'-terminal exons from yeast artificial chromosomes. In: Genome Research. 1995 ; Vol. 4, No. 6. pp. 322-326.
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