Allshire, R. C. (June 1990) Introduction of large linear minichromosomes into Schizosaccharomyces pombe by an improved transformation procedure. Proceedings of the National Academy of Sciences of the United States of America, 87 (11). pp. 4043-4047. ISSN 0027-8424
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Abstract
The efficiency of transformation of Schizosaccharomyces pombe has been increased 10- to 50-fold over previously reported methods. By using 1 microgram of plasmid, 7.0 x 10(5) transformants are regularly obtained. This increased transformation efficiency is mainly due to the inclusion of the cationic liposome-forming reagent Lipofectin in the protocol. Various parameters affecting transformation of Sc. pombe in the presence of Lipofectin have been examined. Lipofectin can also be used to increase transformation efficiency in Saccharomyces cerevisiae. It is also demonstrated that by using this improved transformation procedure, linear minichromosomes of greater than 500 kilobases can be introduced into Sc. pombe with relative ease. These minichromosomes can replicate as stable linear molecules upon reintroduction into Sc. pombe, demonstrating that Sc. pombe telomeres retain function when reintroduced as naked DNA. The ability of Sc. pombe to admit large DNA molecules indicates that it should be feasible to clone large DNA from other organisms in Sc. pombe.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Chromosomes, Fungal/*ultrastructure DNA, Recombinant Electrophoresis, Agar Gel Genetic Vectors Molecular Weight Phosphatidylethanolamines/pharmacology Plasmids Saccharomycetales/*genetics Schizosaccharomyces/*genetics *Transformation, Genetic/drug effects |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function organism description > yeast |
| CSHL Authors: | |
| Communities: | CSHL labs |
| Depositing User: | Matt Covey |
| Date: | June 1990 |
| Date Deposited: | 30 Mar 2016 16:17 |
| Last Modified: | 30 Sep 2019 16:31 |
| PMCID: | PMC54043 |
| Related URLs: | |
| URI: | https://repository.cshl.edu/id/eprint/32304 |
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