Restriction digests of the clone give the following sizes (kb): BamHI--5.8, 0.3; PvuI--6.1; EcoRI--4.0, 2.3. Linearizing with SmaI and transformating into a YAC-containing his3 deletion strain will target integration to Alu sequences. Linearizating at the PvuI site will target integration to the TRP1 centric arm of pYAC4 derivatives. His+ transformants can then be transferred to a eukaryotic cell by spheroplast fusion. YI-type shuttle integrative vector useful for incorporating a neoR cassette into YACs. Constructed by ligating the 1.35 kb XhoI fragment of pPOL2shortneobpA to the largest PvuII fragment of pBP47. The order of the major features in this plasmid is: pMB1 ori - bla - HIS3 - BamHI - SmaI - BamHI - pol2sneo. |
