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Related Experiment Videos

Construction of a genetically engineered strain producing propionylspiramycin.

Y Li1, B Liu, X Li

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

Chinese Journal of Biotechnology
|January 1, 1992
PubMed
Summary

Genetic engineering successfully introduced the propionyl acylase gene into Spiramycin producer S. spiramyceticus. The resulting transformant strain produces propionylspiramycin, a valuable antibiotic compound.

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Biotechnology

Background:

  • Cloning of the propionyl acylase gene and expression in S. mycarofaciens mutant.
  • Previous work established gene cloning and expression in S. lividans TK54.

Purpose of the Study:

  • To transform the pIJM9 recombinant plasmid containing the propionyl acylase gene into S. spiramyceticus.
  • To confirm the successful integration and expression of the gene in the spiramycin producer.

Main Methods:

  • Transformation of S. spiramyceticus with pIJM9 recombinant plasmid.
  • Colony hybridization and Southern hybridization for plasmid confirmation.
  • Thin-layer chromatography (TLC), bioautography, High-Performance Liquid Chromatography (HPLC), and Mass Spectrometry for product analysis.

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Main Results:

  • No. 61 transformant confirmed to harbor the pIJM9 recombinant plasmid via hybridization techniques.
  • TLC and bioautography indicated a component similar to propionylspiramycin.
  • HPLC and Mass Spectrometry confirmed the presence of propionylspiramycin II in fermentation products.

Conclusions:

  • The No. 61 transformant is a genetically engineered strain.
  • This engineered strain is capable of producing propionylspiramycin.