Quantitation of apolipoprotein ε gene expression by competitive polymerase chain reaction in a patient with familial apolipoprotein E deficiency

• Verfasst von: Dobmeyer, Jürgen [VerfasserIn]
• Verfasst von: Rexin, Martin [VerfasserIn]
• Verfasst von: Dobmeyer, Thomas S [VerfasserIn]
• Verfasst von: Klein, Stefan A [VerfasserIn]
• Verfasst von: Rossol, Rita [VerfasserIn]
• Verfasst von: Feussner, Giso [VerfasserIn]
• Titel: Quantitation of apolipoprotein ε gene expression by competitive polymerase chain reaction in a patient with familial apolipoprotein E deficiency
• Verf.angabe: Jürgen M. Dobmeyer, Martin Rexin, Thomas S. Dobmeyer, Stefan A. Klein, Rita Rossol, Giso Feussner
• E-Jahr: 1998
• Jahr: 24 June 1998
• Umfang: 9 S.
• Fussnoten: Gesehen am 30.01.2025
• Titel Quelle: Enthalten in: Clinica chimica acta
• Ort Quelle: Amsterdam [u.a.] : Elsevier Science, 1956
• Jahr Quelle: 1998
• Band/Heft Quelle: 274(1998), 2, Seite 119-137
• ISSN Quelle: 1873-3492
• DOI: doi:10.1016/S0009-8981(98)00046-1
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: Apolipoprotein E
• Sach-SW: Atherosclerosis
• Sach-SW: Gene expression
• Sach-SW: Macrophage
• Sach-SW: Polymerase chain reaction
• K10plus-PPN: 1916026664

Abstract

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.