Premium
Cloning and characterization of the Yarrowia lipolytica squalene synthase ( SQS 1) gene and functional complementation of the Saccharomyces cerevisiae erg 9 mutation
Author(s) -
Merkulov Sergei,
van Assema Friso,
Springer Jan,
Fernandez del Carmen Asun,
Mooibroek H.
Publication year - 2000
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/(sici)1097-0061(200002)16:3<197::aid-yea513>3.0.co;2-l
Subject(s) - biology , complementation , ura3 , yarrowia , saccharomyces cerevisiae , biochemistry , farnesyl diphosphate synthase , gene , farnesyl diphosphate farnesyltransferase , microbiology and biotechnology , genetics , atp synthase , mutant , farnesyltransferase , enzyme , prenylation
The squalene synthase (SQS) gene encodes a key regulatory enzyme, farnesyl‐diphosphate farnesyltransferase (EC 2.5.1.21), in sterol biosynthesis. The SQS 1 gene was isolated from a subgenomic library of the industrially important yeast Yarrowia lipolytica , using PCR‐generated probes. Probes were based on conserved regions of homologues from different organisms. The complete nucleotide sequence of the coding region and the corresponding amino acid sequence were determined. The sequences showed extensive homologies with squalene synthase genes and enzymes from a number of other organisms and extreme amino acid conservation within the binding and catalytic domains. Direct cloning of a 4.3 kb genomic Y. lipolytica fragment, also comprising its own promoter and terminator sequences, into autonomously replicating plasmid YEp352 and subsequent transformation of a Saccharomyces cerevisiae mutant strain with relevant erg 9: ura 3–1 markers, resulted in functional complementation of these deficiencies, although Northern blot analyses did not reveal a unique full‐length messenger. The availability of the Y. lipolytica SQS 1 gene (GenBank Accession No. AF092497) offers prospects for metabolic engineering of the isoprenoid and sterol biosynthetic pathways. Copyright © 2000 John Wiley & Sons, Ltd.