Stanovení exprese genu PML-RARalpha na myším modelu akutní promyelocytární leukemie metodou RT-qPCR za použití RNA izolované z krve

Autor: Švec, David
Jazyk: čeština
Rok vydání: 2007
Druh dokumentu: masterThesis
Popis: Our goal was to develop an accurate, quick and affordable method for following up the progression of acute promyelocytic leucemia in murine model. We took advantage of a developed system of reverse transkriptase quantitative polymerase chain reaction (RT-qPCR) performinng human bone marrow samples to develop a quantitative method of measurement of PML-RARalpha transcripts for murine model. PML-RARalpha transcripts are hallmarks of acute promyelocytic leucemia (APL) with t(15;17) translocation. For measuring bcr1 transcripts, RNA was isolated from murine blood. We present here a method for quantification of t(15;17) transcript bcr1, which is developed in myeloid cells of transgenic mice and then is injected into FVB/N mice. The expression of PML-RARalpha transcripts is normalized using the housekeeping 18S ribosomal subunit gene. We have quantitative data for three healthy mice (FVB/N) and for six ill mice (APL), which were injected 104, 103 or 102 blasts. We show negative expression in FVB/N mice and different copy numbers for APL mice. There is not shown a complete correlation between the amount of injected cells and the amount of transkript copies.
Databáze: Networked Digital Library of Theses & Dissertations
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