Quantitation of apolipoprotein ε gene expression by competitive polymerase chain reaction in a patient with familial apolipoprotein E deficiency
A simple method of obtaining semiquantitative and reliable data on apolipoprotein (apo) ε gene expression is described. We detected apo ε specific sequences by reverse transcription (rT)-PCR. For quantitative measurement, an apo ε DNA standard was produced allowing the development of a competitive P...
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| Main Authors: | , , , , , |
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| Format: | Article (Journal) |
| Language: | English |
| Published: |
24 June 1998
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| In: |
Clinica chimica acta
Year: 1998, Volume: 274, Issue: 2, Pages: 119-137 |
| ISSN: | 1873-3492 |
| DOI: | 10.1016/S0009-8981(98)00046-1 |
| Online Access: | Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1016/S0009-8981(98)00046-1 Verlag, lizenzpflichtig, Volltext: https://www.sciencedirect.com/science/article/pii/S0009898198000461 
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| Author Notes: | Jürgen M. Dobmeyer, Martin Rexin, Thomas S. Dobmeyer, Stefan A. Klein, Rita Rossol, Giso Feussner |
| Summary: | A simple method of obtaining semiquantitative and reliable data on apolipoprotein (apo) ε gene expression is described. We detected apo ε specific sequences by reverse transcription (rT)-PCR. For quantitative measurement, an apo ε DNA standard was produced allowing the development of a competitive PCR-method. The efficiency of RNA extraction and cDNA synthesis was controlled by quantitation of a housekeeping gene (glyceraldehyde-3-phosphatedehydrogenase, G3PDH) in separate reactions. To imitate a defined induction of apo ε gene expression, serial twofold dilutions of total RNA were reversely transcribed and the respective cDNAs used to perform a competitive apo ε and G3PDH PCR. The change in apo ε cDNA and G3PDH cDNA was 1.7-2.3-fold with an expected value of 2.0-fold. Standard deviations in three independently performed experiments were within a range of <15% of the mean, indicating low intra-assay variation and high reproducibility. To illustrate this method, apo ε gene expression was measured in a patient with complete lack of functional active apo E in comparison to healthy controls. The method presented here might be valuable in assessment of apo ε gene expression in human disease. |
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| Item Description: | Gesehen am 30.01.2025 |
| Physical Description: | Online Resource |
| ISSN: | 1873-3492 |
| DOI: | 10.1016/S0009-8981(98)00046-1 |