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Molecular Biology Reagents> | HEK293T cells were seeded on coverslips within the wells of a 6-well plate at a density of 2 ×105 cells/well (for confocal experiments) or in a T75 flask at a density of 3 ×106 cells/flask (for western blot); cells were transiently transfected with 0.5 μg of DNA/well and 8 μg of DNA/flask of each DNA plasmid pcDNA3.1 (EV), pcDNA 3.1 ORF IV (CP), and pcDNA 3.1 ORF VI (TAV) (Genscript) with Polyethylenimine (PEI) 1 : 6 ratio with DNA. | Get A Quote |
Recent comparisons between plant and animal viruses reveal many common principles that underlie how all viruses express their genetic material, amplify their genomes, and link virion assembly with replication. Cauliflower mosaic virus (CaMV) is not infectious for human beings. Here, we show that CaMV transactivator/viroplasmin protein (TAV) shares sequence similarity with and behaves like the human ribonuclease H1 (RNase H1) in reducing DNA/RNA hybrids detected with S9.6 antibody in HEK293T cells. We showed that TAV is clearly expressed in the cytosol and in the nuclei of transiently transfected human cells, similar to its distribution in plants. TAV also showed remarkable cytotoxic effect... More