REP3-derived yeast shuttle vector.
A yeast-shuttle vector using a XbaI-PstI fragment of the 2 microns DNA 1293 bp in length has been constructed. This sequence spans the REP 3 locus and the origin of replication. Besides the 2 microns DNA derived sequences the resulting yeast shuttle vector contains the yeast LEU 2 gene and the Tet' gene of pBR 322. The results demonstrate that the XbaI-PstI fragment is sufficient for proper amplification and partitioning of 2 microns DNA derived yeast-shuttle vectors in yeast. The interruption of the FRT site in this type of vector seems to prevent any recombination between endogenous 2 microns DNA and hybrid plasmid molecules. Moreover, the disturbance of the FLP-system has no influence on the copy number of the plasmid stability.
Schubert
U
U
Central Institute of Microbiology and Experimental Therapy, Academy of Sciences of the GDR, Jena.
Gniel
D
D
Lang
H
H
D015139:Blotting, Southern; D004261:DNA Replication; D004271:DNA, Fungal / Q000302:isolation & purification; D003720:Densitometry; D005784:Gene Amplification; D005822:Genetic Vectors; D008938:Mitosis; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000254:growth & development; D014170:Transformation, Genetic
D004271:DNA, Fungal