Evidence ThatTSG101Aberrant Transcripts Are PCR Artifacts
| Autor: | M. Hampl, H.K. Schackert, Hans-Detlev Saeger, J. Hampl, Jens Plaschke, Gabriele Schackert, Guido Fitze |
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| Rok vydání: | 1998 |
| Předmět: |
Transcription
Genetic Molecular Sequence Data Biophysics Neuroectodermal Tumors Breast Neoplasms macromolecular substances Biology Polymerase Chain Reaction Biochemistry DNA-binding protein law.invention Neuroblastoma Exon Reference Values law Complementary DNA Humans TSG101 Neoplasm Invasiveness splice Neoplasm Metastasis Molecular Biology Polymerase chain reaction Sequence Deletion Genetics Base Composition Leucine Zippers Base Sequence Endosomal Sorting Complexes Required for Transport TSG101 Gene Brain Neoplasms Breakpoint Cell Biology DNA-Binding Proteins Blotting Southern Colonic Neoplasms Female Artifacts Transcription Factors |
| Zdroj: | Biochemical and Biophysical Research Communications. 248:753-760 |
| ISSN: | 0006-291X |
| DOI: | 10.1006/bbrc.1998.9038 |
| Popis: | Critical analysis of the data published so far concerning the TSG101 gene revealed some inconsistencies leading us to its re-evaluation in 80 breast, brain, colon, and neuroectodermal tumors and 37 normal tissue specimens. In this study, the occurrence of TSG101 cDNA aberrant transcripts was verified, but in addition we made observations that are in apparent conflict with the aberrant splicing theory supposed as the underlying mechanism for transcript formation: the location of most deletion breakpoints within exons and nonconformity of these putative splice sites with the highly conserved GT-AG rule, detection of insertions as well as nonreproducible and highly variable results in repeated RT-nested PCRs. Furthermore, we found that reamplification of full-length TSG101 cDNA products leads to the generation of deleted transcripts. In summary, for the first time we provide evidence that the acquired TSG101 transcripts are caused by PCR artifacts. |
| Databáze: | OpenAIRE |
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