Identification of genes regulated by ABF-1 using real time PCR
Poster Number
11
Format
Poster Presentation
Abstract/Artist Statement
Activated B-Cell Factor-1 (ABF-1) has been seen to be a very important repressor in the regulation of the expression of numerous genes in B-cells and muscle cells. In cells infected with the Epstein-Barr Virus (EBV), ABF-1 expression levels have been observed to be above normal. It can be speculated that EBV is using ABF-1 as a defense to repress the expression of other genes that possibly contribute to the resistance of EBV. We have attempted to identify the various genes regulated by ABF-1 using Real Time Polymerase Chain Reaction (Real Time PCR). Real Time PCR is an extremely valuable tool that is used to quantify mRNA levels. B-cells and mouse muscle cells were transfected with ABF-1 in a TREX plasmid. As a control, we transfected B-cells and mouse muscle cells with a TREX plasmid without the ABF-1 gene. The mRNA used in Real Time PCR was obtained from these transfected cells. mRNA isolated from the transfected cells were reverse transcribed into cDNA as the template for Real Time PCR. To amplify our genes of interested we designed primers based on sequences obtained from the data base. Real Time PCR can quantify the DNA by SYBR Green Fluorescence. The data from Real Time PCR of ABF-1 transfected B-cells and control TREX transfected B-cells were compared to look at the regulation of ABF-1 on the various genes of interest. Throughout these studies, we can come to a better understanding of the function of ABF-1 and ultimately find correlation with the interest of EBV in ABF-1.
Location
Pacific Geosciences Center
Start Date
26-4-2003 9:00 AM
End Date
26-4-2003 5:00 PM
Identification of genes regulated by ABF-1 using real time PCR
Pacific Geosciences Center
Activated B-Cell Factor-1 (ABF-1) has been seen to be a very important repressor in the regulation of the expression of numerous genes in B-cells and muscle cells. In cells infected with the Epstein-Barr Virus (EBV), ABF-1 expression levels have been observed to be above normal. It can be speculated that EBV is using ABF-1 as a defense to repress the expression of other genes that possibly contribute to the resistance of EBV. We have attempted to identify the various genes regulated by ABF-1 using Real Time Polymerase Chain Reaction (Real Time PCR). Real Time PCR is an extremely valuable tool that is used to quantify mRNA levels. B-cells and mouse muscle cells were transfected with ABF-1 in a TREX plasmid. As a control, we transfected B-cells and mouse muscle cells with a TREX plasmid without the ABF-1 gene. The mRNA used in Real Time PCR was obtained from these transfected cells. mRNA isolated from the transfected cells were reverse transcribed into cDNA as the template for Real Time PCR. To amplify our genes of interested we designed primers based on sequences obtained from the data base. Real Time PCR can quantify the DNA by SYBR Green Fluorescence. The data from Real Time PCR of ABF-1 transfected B-cells and control TREX transfected B-cells were compared to look at the regulation of ABF-1 on the various genes of interest. Throughout these studies, we can come to a better understanding of the function of ABF-1 and ultimately find correlation with the interest of EBV in ABF-1.