Molecular cloning of the ADE1 gene of Saccharomyces cerevisiae and stability of the transformants |
| |
Authors: | Kenneth Dimock Allen P James Verner L Seligy |
| |
Institution: | Molecular Genetics Section, Division of Biological Sciences, National Research Council of Canada, Ottawa, Ontario K1A 0R6 Canada Tel. (613) 992-6055 |
| |
Abstract: | Plasmid YEp(ADE1)1a, containing a 2.7-kb Sau3A fragment of Saccharomyces cerevisiae DNA inserted at the BamHI site of the yeast shuttle vector pBTI-1 (Morris et al., 1981), results in high frequency, unstable transformation of ade1 yeast strains. A second plasmid, YRp(ADE1)2, containing adjacent 0.5-kb and 3.0-kb BamHI fragments in pBR322 gave three types of yeast transformants: (1) transformants carrying extrachromosomal copies of the plasmid which indicate the presence of a functional ars sequence, (2) transformants indistinguishable from ade1 strains by hybridization analyis, and (3) a transformant carrying a multimeric form of YRp(ADE1)2. Cells transformed with either of the plasmids are free of the red pigment characteristic of ade1 mutants and indicate potential for direct colour-based selection of yeast transformants using ADE1 plasmids. |
| |
Keywords: | Recombinant DNA bank yeast transformation yeast plasmid vectors λ Charon 30 phage restriction endonuclease maps ampicillin-resistant kb kilobase pairs YPD 1% yeast extract 2% peptone 2% dextrose SD 0 67% yeast nitrogen base 2% dextrose (Sherman et al 1981) |
本文献已被 ScienceDirect 等数据库收录! |
|