Time lapse living cells microscopy for studying CML: Preparation of Abl1- and Bcr/Abl - fluorescent protein vectors using Gateway technology

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POTĚŠILOVÁ Michaela VAŘECHA Miroslav TESAŘOVÁ Lenka KRONTORÁD KOUTNÁ Irena

Rok publikování 2010
Druh Článek ve sborníku
Konference Advances in Molecular and Cancer Biology, 2010
Fakulta / Pracoviště MU

Fakulta informatiky

Citace
Obor Genetika a molekulární biologie
Klíčová slova Gateway cloning ABL1 BCR/ABL expression vectors
Popis Using time-lapse microscopy, protein localization and dynamics can be studied directly in living cells. This may bring significant information necessary to complete understanding of protein function. Aberrant tyrosinkinase Bcr/Abl is the main cause of chronic myelogenous leukemia. Abl1 tyrosinkinase represents its native form. Among others these natural and aberrant kinase differ in cell localization, which will be the subject for subsequent examination in the K562 leukemic cell line or in CD34+ cells from bone marrow. We created a unique set of Bcr/Abl and Abl1 expression vectors tagged with EYFP or RFP sequences for examination of protein pools movements in the living cells. The Gateway cloning system was chosen for its flexibility and robustness. We applied two types of fluorescent protein to be able watch both forms of kinase simultaneously.
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