Rapid amplification of immunoglobulin heavy chain switch (IGHS) translocation breakpoints using long-distance inverse PCR
| Autor: | David Oscier, Takashi Sonoki, Tony G. Willis, E L Karran, Martin J. S. Dyer, Reiner Siebert |
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| Rok vydání: | 2004 |
| Předmět: |
Cancer Research
Lymphoma Derivative chromosome Molecular Sequence Data chemical and pharmacologic phenomena Chromosomal translocation Molecular cloning Biology Polymerase Chain Reaction Translocation Genetic Proto-Oncogene Proteins c-myc immune system diseases Proto-Oncogene Proteins hemic and lymphatic diseases Tumor Cells Cultured Humans Receptor Fibroblast Growth Factor Type 3 Cloning Molecular Southern blot Chromosomes Human Pair 14 Gene Rearrangement Genetics Base Sequence Immunoglobulin mu-Chains Inverse polymerase chain reaction Breakpoint PAX5 Transcription Factor Nuclear Proteins Chromosome Breakage Hematology Protein-Tyrosine Kinases Receptors Fibroblast Growth Factor Molecular biology Immunoglobulin Switch Region DNA-Binding Proteins Repressor Proteins Blotting Southern Oncology Proto-Oncogene Proteins c-bcl-6 Immunoglobulin heavy chain PAX5 Carrier Proteins Immunoglobulin Heavy Chains Genes Switch Transcription Factors |
| Zdroj: | Leukemia. 18:2026-2031 |
| ISSN: | 1476-5551 0887-6924 |
| Popis: | Molecular cloning of immunoglobulin heavy chain (IGH) translocation breakpoints identifies genes of biological importance in the development of normal and malignant B cells. Long-distance inverse PCR (LDI-PCR) was first applied to amplification of IGH gene translocations targeted to the joining (IGHJ) regions. We report here successful amplification of the breakpoint of IGH translocations targeted to switch (IGHS) regions by LDI-PCR. To detect IGHS translocations, Southern blot assays using 5' and 3' switch probes were performed. Illegitimate Smu rearrangements were amplified from the 5' end (5'Smu LDI-PCR) from the alternative derivative chromosome, and those of Sgamma or Salpha were amplified from the 3' end (3'Sgamma or 3'alpha LDI-PCR) from the derivative chromosome 14. Using a combination of these methods, we have succeeded in amplifying IGHS translocation breakpoints involving FGFR3/MMSET on 4p16, BCL6 on 3q27, MYC on 8q24, IRTA1 on 1q21 and PAX5 on 9p13 as well as BCL11A on 2p13 and CCND3 on 6p21. The combination of LDI-PCR for IGHJ and IGHS allows rapid molecular cloning of almost all IGH gene translocation breakpoints. |
| Databáze: | OpenAIRE |
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