Heat shock-regulated production of Escherichia coli beta-galactosidase in Saccharomyces cerevisiae - PubMed (original) (raw)

Comparative Study

Heat shock-regulated production of Escherichia coli beta-galactosidase in Saccharomyces cerevisiae

D B Finkelstein et al. Mol Cell Biol. 1983 Sep.

Abstract

The HSP90 gene of the yeast Saccharomyces cerevisiae encodes a heat shock-inducible protein with an Mr of 90,000 (hsp90) and unknown function. We fused DNA fragments of a known sequence (namely, either end of a 1.4-kilobase EcoRI fragment which contains the S. cerevisiae TRP1 gene) to an EcoRI site within the coding sequence of the HSP90 gene. When these fusions are introduced into S. cerevisiae they direct the synthesis of unique truncated hsp90 proteins. By determining the size and charge of these proteins we were able to deduce the translational reading frame at the (EcoRI) fusion site. This information allowed us to design and construct a well-defined in-frame fusion between the S. cerevisiae HSP90 gene and the Escherichia coli lacZ gene. When this fused gene is introduced into S. cerevisiae on a multicopy plasmid vector, it directs the heat shock-inducible synthesis of a fused protein, which is an enzymatically active beta-galactosidase. Thus, for the first time, it is possible to quantitate the heat shock response in a eucaryotic organism with a simple enzyme assay.

PubMed Disclaimer

References

    1. J Bacteriol. 1978 Jun;134(3):1133-40 - PubMed
    1. Arch Biochem Biophys. 1976 May;174(1):52-65 - PubMed
    1. Cell. 1979 Jun;17(2):241-54 - PubMed
    1. Nature. 1979 Nov 1;282(5734):39-43 - PubMed
    1. Gene. 1980 Jul;10(2):157-66 - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources