효모의 ADE1 유전자의 Cloning 에 대하여

Abstract

YEp24, Yeast-E. coli shuttle vector, carry the URA3 gene as a selection marker and contain the replication origin of 2 ㎛ plasmid which is the endogenous plasmid in Saccharomyces cerevisiae and have two cloning sites i.e. BamHI and SalI. We prepared the total gene bank of Saccharomyces cerevisiae by inserting the Sau3A1 partial digested fragments which was size-fractionated into the BamHI site of the YEp 24. The hybrid plasmid pool were isolated from the total gene bank of Saccharomyces cerevisiae to clone the ADEI gene and by using the SHY 3 as yeast host which carry the adel-101 and ura3-52 marker, we could isolate the yeast transformants carried the ADEI gene on synthetic minimal media dificient Adenine and Uracil by complementation. We determined the restriction map and observed the stability of the recombinant plasmids of three transformants among 15 transformants. We observed the close similarity of the two recombinant plasmids in the restriction pattern and the stable maintenance of the recombinant plasmids in non-selective condition.