Expression of yeast DNA mismatch repair proteins and DNA binding characteristics of the yeast condensin Smc2/Smc4 heterodimer;
This thesis describes initial experiments on two separate projects. The first project involved the expression and purification of the DNA mismatch repair proteins Mlhl and Pmsl. Expression vectors were made to singly and co-express these two proteins in yeast. Unfortunately, the proteins were found to be insoluble and this project was put on hold. The second project involved studying the DNA binding characteristics of the Smc2/4 core subunits of the yeast condensin complex. During these studies, it was found that the relatively low ionic strength buffer used for most published DNA binding assays caused these proteins to precipitate. In a higher salt buffer that maintained solubility, Smc2/4 heterodimer was found to bind preferentially to M13mpl8 single strand DNA. These results are discussed in light of on-going experiments with the condensin complex.