Restriction digests of the clone give the following sizes (kb): BamHI--3.7, 2.7, 0.7, 0.6; EcoRI--6.8, 0.9; PvuI--6.8, 0.9; HindIII--3.9, 1.6, 1.2, 1.0. To convert the host phenotype from HIS3 to ADE2, transform with the PstI+SacI digested vector (4.99 kb) and select for Ade+ transformants. A marker swap (HIS3->ADE2) vector for altering a Saccharomyce s cerevisiae host by a one-step gene disruption. When swapping markers on an episomal plasmid, appropriate phenotype may result from loss of the plasmid unless a second selectable or scorable marker is used to ensure plasmid maintenance. Vector was constructed by replacing an internal MscI-NsiI fragment of HIS3 with a 3.7 kb NotI (blunt)-PstI fragment containing the ADE2 coding sequence. ADE2 and HIS3 are in the opposite orientation. |
