VECTORS ENCODING ALTERNATIVE ANTIBIOTIC-RESISTANCE FOR USE IN THE YEAST 2-HYBRID SYSTEM

We have altered the antibiotic resistance of the reporter plasmids and the pJG4-5 activation-domain and pEG202 DNA binding-domain plasmids u sed in the Brent interaction trap/two-hybrid system. These plasmids we re each previously ampicillin-resistant, resulting in an inefficient p urification of any one plasmid from a yeast strain containing all thre e plasmids that constitute the complete interaction trap. By creating derivatives of each of these plasmids expressing either kanamycin or c hloramphenicol resistance, along with the parent-plasmids, we now have the option to use the interaction trap in yeast with three E. coli di fferentially selectable vectors. This will allow isolation of any one plasmid by purifying all of the interaction trap plasmids from yeast s imultaneously and plating E. coli transformed with the plasmids onto t he appropriate antibiotic plate to select the particular plasmid of in terest.