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Single-molecule Imaging of Gene Regulation In vivo Using Cotranslational Activation by Cleavage (CoTrAC)
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[Studies on midecamycin 4"-O-propionyltransferase gene structure]

X Zhang1, Y Wang

  • 1Institute of Medicinal Biotechnology, Chinese Academy of Medical Science, Beijing.

Wei Sheng Wu Xue Bao = Acta Microbiologica Sinica
|December 1, 1996
PubMed

Insights

The midecamycin propionyltransferase (mpt) gene was cloned and characterized. This gene enables the conversion of spiramycin into 4"-O-propionylspiramycin in Streptomyces species.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Context:

  • Midecamycin propionyltransferase (mpt) gene is crucial for antibiotic modification.
  • Understanding the genetic basis of antibiotic biosynthesis and modification is essential for developing new antimicrobial agents.

Purpose:

  • To clone, characterize, and functionally express the midecamycin propionyltransferase (mpt) gene.
  • To elucidate the genetic and molecular properties of the mpt gene and its product.

Summary:

  • A restriction map of the 8.0 kb BamHI-BamHI DNA fragment containing the mpt gene was constructed.
  • Southern blot analysis localized the mpt gene to a 3.0 kb EcoRI-EcoRI-PstI DNA fragment, which was subsequently cloned into the pWHM3 shuttle vector, yielding pWFPE.
  • Sequence analysis revealed an open reading frame encoding a 388 amino acid protein with high similarity to the CarE product. The gene features a consensus ribosomal binding site and promoter elements, with an inverted repeat downstream serving as a transcriptional terminator.

Impact:

  • The cloned mpt gene, when expressed in *S. ambofaciens* and *S. lividans*, confers the ability to convert spiramycin into 4"-O-propionylspiramycin.
  • This research provides insights into the enzymatic modification of macrolide antibiotics and facilitates genetic engineering of antibiotic production in Streptomyces.

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