Cloning and nucleotide sequence of the ispA gene responsible for farnesyl diphosphate synthase activity in Escherichia coli

J Biochem. 1990 Dec;108(6):995-1000. doi: 10.1093/oxfordjournals.jbchem.a123327.


The molecular cloning and the determination of the nucleotide sequence of the ispA gene responsible for farnesyl diphosphate (FPP) synthase [EC] activity in Escherichia coli are described. E. coli ispA strains have temperature-sensitive FPP synthase, and the defective gene is located at about min 10 on the chromosome. The wild-type ispA gene was subcloned from a lambda phage clone containing the chromosomal fragment around min 10, picked up from the aligned genomic library of Kohara et al. [Kohara, Y., Akiyama, K., & Isono, K. (1987) Cell 50, 495-508]. The cloned gene was identified as the ispA gene by the recovery and amplification of FPP synthase activity in an ispA strain. A 1,452-nucleotide sequence of the cloned fragment was determined. This sequence specifies two open reading frames, ORF-1 and ORF-2, encoding proteins with the expected molecular weights of 8,951 and 32,158, respectively. A part of the deduced amino acid sequence of ORF-2 showed similarity to the sequences of eucaryotic FPP synthases and of crtE product of a photosynthetic bacterium. The plasmid carrying ORF-2 downstream of the lac promoter complemented the defect of FPP synthase activity of the ispA mutant, showing that the product encoded by ORF-2 is the ispA product. The maxicell analysis indicated that a protein of molecular weight 36,000, approximately consistent with the molecular weight of the deduced ORF-2-encoded protein, is the gene product.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkyl and Aryl Transferases*
  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • Codon
  • Escherichia coli / enzymology
  • Escherichia coli / genetics*
  • Geranyltranstransferase
  • Hemiterpenes*
  • Molecular Sequence Data
  • Molecular Weight
  • Open Reading Frames
  • Organophosphorus Compounds / metabolism
  • Restriction Mapping
  • Sequence Homology, Nucleic Acid
  • Temperature
  • Transferases / biosynthesis
  • Transferases / genetics*


  • Codon
  • Hemiterpenes
  • Organophosphorus Compounds
  • isopentenyl pyrophosphate
  • Transferases
  • Alkyl and Aryl Transferases
  • Geranyltranstransferase

Associated data

  • GENBANK/D00694