Introduction of large linear minichromosomes into Schizosaccharomyces pombe by an improved transformation procedure

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

URL: http://www.ncbi.nlm.nih.gov/pubmed/2349217
DOI: 10.1073/pnas.87.11.4043

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 Mar 2016 16:17
PMCID: PMC54043
Related URLs:
URI: http://repository.cshl.edu/id/eprint/32304

Actions (login required)

Administrator's edit/view item Administrator's edit/view item
CSHL HomeAbout CSHLResearchEducationNews & FeaturesCampus & Public EventsCareersGiving