Restriction digests of the clone give the following sizes (kb): EcoRI--15.9; XhoI--11.0, 3.4, 1.0; BamHI--14.0, 1.85; BamHI/EcoRI--8.2, 5.8, 1.85; SalI/BamHI--7.4, 6.6, 1.85. To facilitate chromosome walking, the ends of the insert can be recovered in E. coli. The artificial chromosomal DNA can be digested with XhoI, ligated, and used to transform E. coli to amp or kan resistance. Cloning into the EcoRI site insertionally inactivates SUP4. G418 resistance is expressed in mammalian cell lines to permit genetic complementation experiments. Constructed by Christopher Traver. XhoI linkers were ligated to the blunt ended 4.2 kb PvuI/BamHI fragment of pSV2neo (with neomycin resistance and ColE1 origin). This was then cloned into the SalI site of pYAC4. Contains telomer elements from Tetrahymena. |
