Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 1986 Sep 11;14(17):6871-84.
doi: 10.1093/nar/14.17.6871.

Efficient isolation of the linear DNA killer plasmid of Kluyveromyces lactis: evidence for location and expression in the cytoplasm and characterization of their terminally bound proteins

J C Stam, J Kwakman, M Meijer, A R Stuitje
Free PMC article

Efficient isolation of the linear DNA killer plasmid of Kluyveromyces lactis: evidence for location and expression in the cytoplasm and characterization of their terminally bound proteins

J C Stam et al. Nucleic Acids Res. .
Free PMC article

Abstract

Differential centrifugation of an osmotic lysate of K. lactis protoplasts showed that the linear DNA killer plasmids of K. lactis, pGKL1 and pGKL2, are almost exclusively present in the cytoplasmic fraction. This fractionation procedure allows the rapid isolation of large amounts of plasmid DNA without contamination by chromosomal and mitochondrial DNA. With these DNA preparations the size of the terminally bound proteins was estimated to be 28 and 36 kDal for pGKL1 and pGKL2, respectively. The entire pGKL1 sequence (except for 21 base pairs at the right terminus) was cloned in a shuttle vector that permits autonomous replication in the nucleus of K. lactis. However, killer gene expression could not be established in transformants. In connection with the observed cytoplasmic localization, this result suggests that gene expression of the killer DNA plasmids is entirely cytoplasmic.

PubMed Disclaimer

References

    1. Nucleic Acids Res. 1984 Jul 25;12(14):5685-92 - PubMed
    1. Mol Gen Genet. 1979 Jan 5;168(1):1-25 - PubMed
    1. Nature. 1970 Aug 15;227(5259):680-5 - PubMed
    1. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5105-9 - PubMed
    1. Nucleic Acids Res. 1982 Dec 20;10(24):8181-90 - PubMed