are goal is the isolation and characterization of megakaryocyte- specific gene products in normal and disease states. To this end we plan the following steps: (1) isolation of mRNA from peripheral blood platelets or from megakaryocytes grown in culture from peripheral blood or bone marrow; (2) reverse transcription to make cDNA; (3) a subtraction procedure to enrich for gene products that are only expressed in disease states or only in megakaryocytes; (4) cloning of these specific messages; (5) sequencing and further characterization. Each of these steps involves further technical development. We have successfully isolated mRNA from platelets, and used this mRNA as a template for RT-PCR. Initial attempts to enrich by subtraction have not yet been successful. We have also grown megakaryocytes from peripheral blood mononuclear cells purified over a ficoll gradient, and are currently optimizing methods for confirming the identity of these cells and isolating them at an acceptable purity.

Agency
National Institute of Health (NIH)
Institute
Clinical Center (CLC)
Type
Intramural Research (Z01)
Project #
1Z01CL010238-01
Application #
3752283
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Clinical Center
Department
Type
DUNS #
City
State
Country
United States
Zip Code