Panhandle polymerase chain reaction amplifies MLL genomic translocation breakpoint involving unknown partner gene
| Autor: | C A, Felix, C S, Kim, M D, Megonigal, D J, Slater, D H, Jones, N B, Spinner, T, Stump, M R, Hosler, P C, Nowell, B J, Lange, E F, Rappaport |
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| Rok vydání: | 1998 |
| Předmět: |
Base Sequence
Molecular Sequence Data Infant Histone-Lysine N-Methyltransferase Precursor Cell Lymphoblastic Leukemia-Lymphoma Polymerase Chain Reaction Introns Translocation Genetic DNA-Binding Proteins Proto-Oncogenes Humans Female Chromosomes Human Pair 4 Myeloid-Lymphoid Leukemia Protein Transcription Factors |
| Zdroj: | Blood. 90(12) |
| ISSN: | 0006-4971 |
| Popis: | We used a new approach called panhandle polymerase chain reaction (PCR) to clone an MLL genomic translocation breakpoint in a case of acute lymphoblastic leukemia of infancy in which karyotype analysis was technically unsuccessful and did not show the translocation partner. Panhandle PCR amplified known MLL sequence 5' of the breakpoint and 3' sequence from the unknown partner gene from a DNA template with an intrastrand loop schematically shaped like a pan with a handle. The 7-kb panhandle PCR product contained the translocation breakpoint in MLL intron 8. The partner DNA included unique nonrepetitive sequences, Alu and mammalian apparent LTR-retrotransposon (MaLR) repetitive sequences, and a region of homology to expressed sequence tags. MaLR sequences have not been found before near leukemia-associated translocation breakpoints. The nonrepetitive sequences were not homologous to known partner genes of MLL. Screening of somatic cell hybrid and radiation hybrid lines by PCR and fluorescence in situ hybridization analysis of normal metaphase chromosomes mapped the partner DNA to chromosome band 4q21. Reverse transcriptase-PCR identified an MLL-AF-4 chimeric mRNA, indicating that panhandle PCR identified a fusion of MLL with a previously uncharacterized AF-4 intronic sequence. Panhandle PCR facilitates cloning translocation breakpoints and identifying unknown partner genes. |
| Databáze: | OpenAIRE |
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