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dc.contributor.authorRossiter, Belinda J*
dc.contributor.authorFuscoe, J C*
dc.contributor.authorMuzny, D M*
dc.contributor.authorFox, Margaret*
dc.contributor.authorCaskey, C T*
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.
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
html.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.


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