Tax and TORC 3/HTLV Gene Expression

As part of my research I will be working with 293T cells and analyzing TORC and Tax localization in the cells.   I will also be analyzing the localization of Tax and TORC in 293 cells and than comparing both of the localization in normal vs. cancerous cells.   The main goal of the research is to analyze the gene expression of HTLV in the presence of tax and cellular transcription factors, which are thought to enhance HTLV gene expression.  HTVL is the first retrovirus that was discovered and it is known to develop an often rapidly fatal leukemia, debilitative myelopathy, uveitis, dermatitis, and inflammatory disorder among others.  The goal of this academic year is to finalize our findings and determine if TORC 3 enhances HTLV gene expression.

 

To study localization we will be using immunofluorescence technique.  For this technique we use antibodies that are specific to tax and TORC 3.  Each antibody will emit a specific wavelength of light once excited allowing us to view it under the microscope. The light is specific to the antibody (red or green) and is only emitted when the antibody is present in the cells, indicating the presence of the protein of interest in the cells.   Prior to staining, we will transfect cells with Tax and TORC, stain with the antibody, and view the slides under a special microscope.  To transfect cells we must grow them in a special media and incubate them in a regulated environment to ensure their growth occurs at optimal conditions.   Many times many trials of immunofluorescence are needed to obtain pictures that are of good quality. I look forward to all the exciting new challenges this academic year will bring.

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