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The expression, localization, and effect of a human interferon in plants
G A De Zoeten1, J R Penswick, M A Horisberger
1Friederich Miescher-Institut, Basel, Switzerland.
Virology
|September 1, 1989
Summary
Researchers engineered Cauliflower mosaic virus (CaMV) to produce human interferon alpha D (IFN alpha D) in plants. The plant-produced interferon was biologically active and did not impede viral superinfection.
Area of Science:
- Plant Virology
- Molecular Biology
- Biotechnology
Background:
- Cauliflower mosaic virus (CaMV) is a plant DNA virus used as a vector in plant biotechnology.
- Human interferon alpha D (IFN alpha D) is a protein with antiviral properties.
Purpose of the Study:
- To engineer a stable CaMV strain (Ca524i) capable of expressing biologically active human IFN alpha D in plants.
- To investigate the biological activity and localization of plant-expressed IFN alpha D.
Main Methods:
- Genetic modification of CaMV ORF II to insert the human IFN alpha D coding sequence.
- Inoculation of turnip plants (Brassica rapa) with modified CaMV DNA.
- Assay of IFN alpha D biological activity in infected plants.
- Electron microscopy and antibody gold labeling to determine IFN localization.
Main Results:
- A stable CaMV strain (Ca524i) successfully produced biologically active human IFN alpha D in infected turnip plants.
- A mutant strain (Ca562i) with a deleted CYS codon also produced active IFN alpha D.
- Plant-produced IFN alpha D did not inhibit superinfection by turnip yellow mosaic virus (TYMV).
- IFN alpha D was localized within CaMV inclusion bodies in infected plant cells.
Conclusions:
- CaMV can be effectively engineered to express functional human IFN alpha D in planta.
- The expressed IFN alpha D is biologically active but does not confer broad antiviral resistance against unrelated viruses like TYMV.
- CaMV inclusion bodies serve as a localization site for the expressed interferon within infected plant cells.