title: Expression of the Saccharomyces cerevisiae PIS gene and synthesis of phosphatidylinositol in Escherichia coli.
abstract: Expression of the Saccharomyces cerevisiae PIS gene encoding phosphatidylinositol synthase in Escherichia coli was achieved by inserting its coding sequence into lacZ on pUC8. The fused gene encoded a phosphatidylinositol synthase whose amino-terminal three amino acids had been replaced by the amino-terminal five amino acids of E. coli beta-galactosidase. E. coli cells bearing this recombinant plasmid produced a significant level of phosphatidylinositol synthase in the presence of a lacZ inducer, isopropylthio-beta-D-galactopyranoside. When the culture medium was supplemented with myo-inositol and isopropylthio-beta-D-galactopyranoside, the cells accumulated a substantial amount of phosphatidylinositol in their membranes. When a saturating level of myo-inositol was added, phosphatidylinositol constituted about 4% of the total phospholipids. Phosphatidylinositol accumulation occurred at the expense of phosphatidylglycerol. The ratio of phosphatidylethanolamine to total acidic phospholipids remained constant. The growth rate of phosphatidylinositol-containing E. coli cells did not differ significantly from that of cells with the normal phospholipid composition.
authors.affiliation: Department of Biochemistry, Gunma University School of Medicine, Maebashi, Japan.
authors.forename: J
authors.initials: J
authors.lastname: Nikawa
authors.forename: T
authors.initials: T
authors.lastname: Kodaki
authors.forename: S
authors.initials: S
authors.lastname: Yamashita
mesh_terms: D051607:CDP-Diacylglycerol-Inositol 3-Phosphatidyltransferase; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D005786:Gene Expression Regulation; D008563:Membrane Lipids / Q000096:biosynthesis; D010715:Phosphatidylglycerols / Q000096:biosynthesis; D010716:Phosphatidylinositols / Q000096:biosynthesis*; D010770:Phosphotransferases / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D017855:Transferases (Other Substituted Phosphate Groups)
chemical_list: D008563:Membrane Lipids; D010715:Phosphatidylglycerols; D010716:Phosphatidylinositols; D010770:Phosphotransferases; D017855:Transferases (Other Substituted Phosphate Groups); D051607:CDP-Diacylglycerol-Inositol 3-Phosphatidyltransferase