PMID: 8604345Mar 15, 1996Paper

Scanning for genes in large genomic regions: cosmid-based exon trapping of multiple exons in a single product

Nucleic Acids Research
N A DatsonJ T Den Dunnen

Abstract

To facilitate the scanning of large genomic regions for the presence of exonic gene segments we have constructed a cosmid-based exon trap vector. The vector serves a dual purpose since it is also suitable for contig construction and physical mapping. The exon trap cassette of vector sCOGH1 consists of the human growth hormone gene driven by the mouse mettallothionein-1 promoter. Inserts are cloned in the multicloning site located in intron 2 of the hGH gene. The efficiency of the system is demonstrated with cosmids containing multiple exons of the Duchenne Muscular Dystrophy gene. All exons present in the inserts were successfully retrieved and no cryptic products were detected. Up to seven exons were isolated simultaneously in a single spliced product. The system has been extended by a transcription-translation-test protocol to determine the presence of large open reading frames in the trapped products, using a combination of tailed PCR primers directing protein synthesis in three different reading frames, followed by in vitro transcription-translation. Having larger stretches of coding sequence in a single exon trap product rather than small single exons greatly facilitates further analysis of potential genes and offers new p...Continue Reading

References

Apr 1, 1992·Human Molecular Genetics·J T Den DunnenH Y Steensma
May 1, 1991·Proceedings of the National Academy of Sciences of the United States of America·A J BucklerD E Housman
Nov 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·G M DuykD R Cox
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Oct 11, 1994·Nucleic Acids Research·N A DatsonJ T Den Dunnen
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Nov 11, 1993·Nucleic Acids Research·D B Krizman, S M Berget
Oct 1, 1993·Human Molecular Genetics·P A RoestJ T den Dunnen

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Citations

Jun 13, 1998·Genes, Chromosomes & Cancer·J AdélaïdeM J Pébusque
Aug 6, 1999·Gene·E M van SchothorstP Devilee
Aug 7, 1999·Insect Biochemistry and Molecular Biology·S E BrownD L Knudson
Sep 1, 1997·Nature Genetics·UNKNOWN French FMF Consortium
Jul 2, 1999·Journal of Sleep Research·U Schibler, M Tafti
Jul 1, 1997·Mammalian Genome : Official Journal of the International Mammalian Genome Society·E Van de VosseJ T Den Dunnen
Feb 9, 2000·Genomics·P CousinP Shaw
May 12, 2016·European Journal of Human Genetics : EJHG·Johannes G DauwerseDorien Peters

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