Abstract
A newE. coli — S. cerevisiae shuttle plasmid cloning vector (pPW263) with a positive type of selection, was constructed. The selection system, based on the regulatory region of λ phage controlling the expression of tetracycline resistance, was derived from the cloning vector pUN121 (Nilssonet al. 1983). There are three cloning sites in thecI gene,EcoRI,HindIII andBglll, and, in addition, two unique sites in the neighborhood,BamHI andSalI. The size of the vector is 7.8 kb. The maintenance of the vector and the selection in yeast was ensured by the replication region of the 2μ plasmid and by theURA3 marker gene, respectively.
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Abbreviations
- ApR :
-
ampicillin resistance
- cat:
-
chloramphenicol-acetyl transferase gene
- cI:
-
λ repressor gene
- KmR :
-
gene coding for resistance to kanamycin
- LiAc:
-
lithium acetate
- oRPR :
-
right operator-promoter region of λ phage
- TcR :
-
tetracycline resistance
- TcI R :
-
coding region of tetracycline resistance gene
- URA3:
-
gene ofS. cerevisiae coding for orotidine-5′-phosphate decarboxylase
- 2μ:
-
2μm plasmid sequence
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Půta, F., Wambutt, R. Construction of a newEscherichia coli — Saccharomyces cerevisiae shuttle plasmid cloning vector allowing positive selection for cloned fragments. Folia Microbiol 37, 193–198 (1992). https://doi.org/10.1007/BF02933146
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DOI: https://doi.org/10.1007/BF02933146