Decorating dsDNA with a functional belt : bisAGT zif-finger protein for site-specific tagging of DNA.

Abstract

Being the best understood DNA binding proteins, Zinc finger proteins can recognize and discriminate closely related sequences in both vitro and vivo (Klug, 2010). This ability has made them to be a promising candidate in creating a system that can detect DNAs in their native forms. In more than two years, the group of Professor Johnsson, EPFL, Switzerland, has been trying to develop a fusion protein between Zif 268, a Zinc finger protein, and two Snap-tags, which are useful self-labeling tool. Such protein, when catenating with a Zif-target containing plasmid, is believed to have many good biological applications. Unfortunately, the successful catenation reaction was observed to have low efficiency and selectivity due to dimensional problems. In this project, we aimed to introduce two elastic unstructured linkers to two locations between Zif268 and each Snap-tag to overcome such hindrance matter. A new protein with linker was highly expressed; however, its catenation efficiency seemed to be unchanged.