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dc.contributor.authorHeighway, Jim
dc.contributor.authorBetticher, Daniel C
dc.contributor.authorHoban, Paul R
dc.contributor.authorAltermatt, H J
dc.contributor.authorCowen, Rachel L
dc.date.accessioned2010-04-06T08:50:54Z
dc.date.available2010-04-06T08:50:54Z
dc.date.issued1996-07-01
dc.identifier.citationCoamplification in tumors of KRAS2, type 2 inositol 1,4,5 triphosphate receptor gene, and a novel human gene, KRAG. 1996, 35 (1):207-14 Genomicsen
dc.identifier.issn0888-7543
dc.identifier.pmid8661122
dc.identifier.doi10.1006/geno.1996.0340
dc.identifier.urihttp://hdl.handle.net/10541/95595
dc.description.abstractAnalysis of a region of DNA, coamplified in tumors with KRAS2, resulted in the identification of the human homologue of the mouse KRAG gene. The gene was widely expressed in a range of cell lines, tumors, and normal tissue and demonstrated a high degree of alternate splicing. A human KRAG cDNA sequence, with a structure similar to that encoded by the amplified gene in mouse Y1 adrenal carcinoma cells, was isolated by RT-PCR. The predicted amino acid similarity between the two sequences was 91%, and hydrophobicity plots suggested a structure closely resembling that of transmembrane 4 superfamily members. Identification of a PCR-based restriction fragment length polymorphism confirmed biallelic expression of KRAG but suggested allele-specific splicing differences in tumors. Northern analysis of mRNA derived from a range of tissues suggested high level expression in muscle and confirmed alternate splicing. To facilitate the analysis of exon junctions, a YAC clone encoding the genomic sequence was identified. This allowed the localization of KRAG to human chromosome 12p11.2. Isolation of one end of this nonchimeric clone demonstrated a perfect match with a 247-bp sequence within the 3' untranslated region of the type 2 1,4, 5-inositol triphosphate receptor gene. Multiplex PCR confirmed the inclusion of both genes in the KRAS2 amplicon in human malignancy, suggesting that either may contribute to the malignant phenotype.
dc.language.isoenen
dc.subjectCanceren
dc.subjectCancer DNAen
dc.subjectCancer Proteinsen
dc.subjectCancer RNAen
dc.subjectLung Canceren
dc.subjectCultured Tumour Cellsen
dc.subject.meshAlleles
dc.subject.meshAmino Acid Sequence
dc.subject.meshAnimals
dc.subject.meshBase Sequence
dc.subject.meshCalcium Channels
dc.subject.meshCarcinoma, Non-Small-Cell Lung
dc.subject.meshCarcinoma, Squamous Cell
dc.subject.meshCarrier Proteins
dc.subject.meshCell Transformation, Neoplastic
dc.subject.meshChromosomes, Human, Pair 12
dc.subject.meshCloning, Molecular
dc.subject.meshDNA, Complementary
dc.subject.meshDNA, Neoplasm
dc.subject.meshGene Amplification
dc.subject.meshGene Expression Regulation, Neoplastic
dc.subject.meshGenes, ras
dc.subject.meshHumans
dc.subject.meshInositol 1,4,5-Trisphosphate Receptors
dc.subject.meshLung Neoplasms
dc.subject.meshMembrane Proteins
dc.subject.meshMice
dc.subject.meshMolecular Sequence Data
dc.subject.meshNeoplasm Proteins
dc.subject.meshNeoplasms
dc.subject.meshRNA Splicing
dc.subject.meshRNA, Messenger
dc.subject.meshRNA, Neoplasm
dc.subject.meshReceptors, Cytoplasmic and Nuclear
dc.subject.meshSequence Alignment
dc.subject.meshSequence Homology, Amino Acid
dc.subject.meshSpecies Specificity
dc.subject.meshTumor Cells, Cultured
dc.titleCoamplification in tumors of KRAS2, type 2 inositol 1,4,5 triphosphate receptor gene, and a novel human gene, KRAG.en
dc.typeArticleen
dc.contributor.departmentCRC Department of Cancer Genetics, Paterson Institute for Cancer Research, Christie Hospital (NHS) Trust, Wilmslow Road, Manchester, M20 9BX, United Kingdom.en
dc.identifier.journalGenomicsen
html.description.abstractAnalysis of a region of DNA, coamplified in tumors with KRAS2, resulted in the identification of the human homologue of the mouse KRAG gene. The gene was widely expressed in a range of cell lines, tumors, and normal tissue and demonstrated a high degree of alternate splicing. A human KRAG cDNA sequence, with a structure similar to that encoded by the amplified gene in mouse Y1 adrenal carcinoma cells, was isolated by RT-PCR. The predicted amino acid similarity between the two sequences was 91%, and hydrophobicity plots suggested a structure closely resembling that of transmembrane 4 superfamily members. Identification of a PCR-based restriction fragment length polymorphism confirmed biallelic expression of KRAG but suggested allele-specific splicing differences in tumors. Northern analysis of mRNA derived from a range of tissues suggested high level expression in muscle and confirmed alternate splicing. To facilitate the analysis of exon junctions, a YAC clone encoding the genomic sequence was identified. This allowed the localization of KRAG to human chromosome 12p11.2. Isolation of one end of this nonchimeric clone demonstrated a perfect match with a 247-bp sequence within the 3' untranslated region of the type 2 1,4, 5-inositol triphosphate receptor gene. Multiplex PCR confirmed the inclusion of both genes in the KRAS2 amplicon in human malignancy, suggesting that either may contribute to the malignant phenotype.


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