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Proteins, Expression, Isolation and Analysis | … The full-length Tag and Tag(84–130) proteins used in isothermal titration calorimetry (ITC) and nuclear magnetic resonance (NMR) studies were treated with H3C PreScission protease (Genscript, Piscataway, NJ) and the cleaved tags were removed by passage through a … | Get A Quote |
Simian virus 40 (SV40) serves as an important model organism for studying eukaryotic DNA replication. Its helicase, Large T-antigen (Tag), is a multi-functional protein that interacts with multiple host proteins, including the ubiquitous ssDNA binding protein Replication Protein A (RPA). Tag recruits RPA, actively loads it onto the unwound DNA, and together they promote priming of the template. Although interactions of Tag with RPA have been mapped, no interaction between Tag and the N-terminal protein interaction domain of the RPA 70kDa subunit (RPA70N) has been reported. Here we provide evidence of direct physical interaction of Tag with RPA70N and map the binding sites using a series of pull-down... More