Time lapse living cells microscopy for studying CML: Preparation of Abl1- and Bcr/Abl - fluorescent protein vectors using Gateway technology
| Authors | |
|---|---|
| Year of publication | 2010 |
| Type | Article in Proceedings |
| Conference | Advances in Molecular and Cancer Biology, 2010 |
| MU Faculty or unit | |
| Citation | |
| Field | Genetics and molecular biology |
| Keywords | Gateway cloning ABL1 BCR/ABL expression vectors |
| Description | 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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