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Related Experiment Videos

Combinatorial natural products: from cloning to analysis.

Paul Budworth1, Julia Khandurina, Andras Guttman

  • 1Horváth Laboratory of Bioseparation Science, University of Innsbruck, Innsbruck, Austria.

Current Medicinal Chemistry
|March 26, 2005
PubMed
Summary

Researchers utilized bioinformatics and genetic engineering to modify plant biosynthetic pathways, successfully over-expressing enzymes in tobacco cell cultures to generate novel medicinal compounds.

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Area of Science:

  • Plant biochemistry and metabolic engineering
  • Bioinformatics and genetic technology applications

Background:

  • Plant secondary metabolites are a rich source of medicinal compounds.
  • Bioinformatics and genetic technologies enable manipulation of plant biosynthetic pathways.
  • Novel chemical entities can be generated by altering these pathways.

Purpose of the Study:

  • To identify and isolate plant genes involved in secondary metabolite biosynthesis.
  • To over-express these genes in a heterologous host system (tobacco cell cultures).
  • To analyze the resulting changes in metabolite profiles.

Main Methods:

  • Interrogation of public databases for plant secondary metabolite enzyme genes.
  • Cloning and transfection of identified genes into tobacco cell cultures using viral vectors.

Related Experiment Videos

  • Over-expression of biosynthetic enzymes in transfected cells.
  • Automated solvent extraction of metabolites from plant tissues and cell cultures.
  • Analysis of metabolite composition using High-Performance Liquid Chromatography (HPLC) and Capillary Electrophoresis (CE).

    • Main Results:

    • Successful identification and isolation of relevant plant genes.
    • Demonstrated over-expression of plant biosynthetic enzymes in tobacco cell cultures.
    • Detected alterations in secondary metabolite patterns in transfected cells compared to controls.
    • Validated the use of HPLC and CE for monitoring these metabolic changes.

    Conclusions:

    • Genetic engineering and bioinformatics provide powerful tools for manipulating plant metabolic pathways.
    • Heterologous expression of plant genes in tobacco cell cultures can lead to the production of novel or modified secondary metabolites.
    • This approach holds potential for the discovery and production of new medicinal compounds from plants.