The Chinese hamster HPRT gene: restriction map, sequence analysis, and multiplex PCR deletion screen.

2.50
Hdl Handle:
http://hdl.handle.net/10541/104697
Title:
The Chinese hamster HPRT gene: restriction map, sequence analysis, and multiplex PCR deletion screen.
Authors:
Rossiter, Belinda J; Fuscoe, J C; Muzny, D M; Fox, Margaret; Caskey, C T
Abstract:
The fine structure of the Chinese hamster hypoxanthine guanine phosphoribosyltransferase (HPRT) gene has been determined; the gene has nine exons and is dispersed over 36 kb DNA. Exons 2-9 are contained within overlapping lambda bacteriophage clones and exon 1 was obtained by an inverse polymerase chain reaction (PCR). All the exons have been sequenced, together with their immediate flanking regions, and these sequences compared to those of the mouse and human HPRT genes. Sequences immediately flanking all exons but the first show considerable homology between the different species but the region around exon 1 is less conserved, apart from the preserved location of putative functional elements. Oligonucleotide primers derived from sequences flanking the HPRT gene exons were used to amplify simultaneously seven exon-containing fragments in a multiplex PCR. This simple procedure was used to identify total and partial gene deletions among Chinese hamster HPRT-deficient mutants. The multiplex PCR is quicker to perform than Southern analysis, traditionally used to study such mutants, and also provides specific exon-containing fragments for further analysis. The Chinese hamster HPRT gene is often used as a target for mutation studies in vitro because of the ease of selection of forward and reverse mutants; the information presented here will enhance the means of investigating molecular defects within this gene.
Affiliation:
Institute of Molecular Genetics, Baylor College of Medicine, Houston, Texas 77030.
Citation:
The Chinese hamster HPRT gene: restriction map, sequence analysis, and multiplex PCR deletion screen. 1991, 9 (2):247-56 Genomics
Journal:
Genomics
Issue Date:
Feb-1991
URI:
http://hdl.handle.net/10541/104697
DOI:
10.1016/0888-7543(91)90249-E
PubMed ID:
2004774
Type:
Article
Language:
en
ISSN:
0888-7543
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorRossiter, Belinda Jen
dc.contributor.authorFuscoe, J Cen
dc.contributor.authorMuzny, D Men
dc.contributor.authorFox, Margareten
dc.contributor.authorCaskey, C Ten
dc.date.accessioned2010-06-11T11:22:18Z-
dc.date.available2010-06-11T11:22:18Z-
dc.date.issued1991-02-
dc.identifier.citationThe Chinese hamster HPRT gene: restriction map, sequence analysis, and multiplex PCR deletion screen. 1991, 9 (2):247-56 Genomicsen
dc.identifier.issn0888-7543-
dc.identifier.pmid2004774-
dc.identifier.doi10.1016/0888-7543(91)90249-E-
dc.identifier.urihttp://hdl.handle.net/10541/104697-
dc.description.abstractThe fine structure of the Chinese hamster hypoxanthine guanine phosphoribosyltransferase (HPRT) gene has been determined; the gene has nine exons and is dispersed over 36 kb DNA. Exons 2-9 are contained within overlapping lambda bacteriophage clones and exon 1 was obtained by an inverse polymerase chain reaction (PCR). All the exons have been sequenced, together with their immediate flanking regions, and these sequences compared to those of the mouse and human HPRT genes. Sequences immediately flanking all exons but the first show considerable homology between the different species but the region around exon 1 is less conserved, apart from the preserved location of putative functional elements. Oligonucleotide primers derived from sequences flanking the HPRT gene exons were used to amplify simultaneously seven exon-containing fragments in a multiplex PCR. This simple procedure was used to identify total and partial gene deletions among Chinese hamster HPRT-deficient mutants. The multiplex PCR is quicker to perform than Southern analysis, traditionally used to study such mutants, and also provides specific exon-containing fragments for further analysis. The Chinese hamster HPRT gene is often used as a target for mutation studies in vitro because of the ease of selection of forward and reverse mutants; the information presented here will enhance the means of investigating molecular defects within this gene.en
dc.language.isoenen
dc.subject.meshAnimals-
dc.subject.meshBase Sequence-
dc.subject.meshBlotting, Southern-
dc.subject.meshCell Line-
dc.subject.meshChromosome Deletion-
dc.subject.meshCloning, Molecular-
dc.subject.meshCricetinae-
dc.subject.meshCricetulus-
dc.subject.meshDNA-
dc.subject.meshExons-
dc.subject.meshHumans-
dc.subject.meshHypoxanthine Phosphoribosyltransferase-
dc.subject.meshMice-
dc.subject.meshMolecular Sequence Data-
dc.subject.meshPolymerase Chain Reaction-
dc.subject.meshRestriction Mapping-
dc.subject.meshSequence Alignment-
dc.subject.meshSequence Homology, Nucleic Acid-
dc.titleThe Chinese hamster HPRT gene: restriction map, sequence analysis, and multiplex PCR deletion screen.en
dc.typeArticleen
dc.contributor.departmentInstitute of Molecular Genetics, Baylor College of Medicine, Houston, Texas 77030.en
dc.identifier.journalGenomicsen
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