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Science and research

Grant Projects (ongoing in the year 2009)

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Development of a Novel Recombinogenic Technique for Chloroplast Transformation and its Use for Production of human papillomavirus E7 Protein in Plants

IAA500960903 [2009 – 2012]

Doc. RNDr. Josef Vlasák, CSc., Biologické centrum AV ČR, v.v.i.
RNDr. Viera Ludvíková

Our aim is to develop more efficient method for plant chloroplast transformation using biolistic bombardment with linear DNA vectors together with chloroplast cassettes expressing bacteriophage lambda Red proteins. The idea stems from experiments with targeted insertion mutagenesis in yeast, cyanobacteria, and E. coli, where DNA ends promote recombination, and lambda Red proteins Exo, Beta, and Gam are very efficient in short recombinogenic ends protection and processing. Linear vectors can be generated conveniently as PCR products with synthetic primer ends that are homologous to sequence targets in the chloroplast genome. Chloroplast vectors will carry marker genes and also human papillomavirus oncogene E7 in the form of highly immunogenic but nononcogenic fusion E7GGG/GUS. It is our goal to obtain transplastomic plants producing this antigen with potential use as therapeutic vaccine. Viral antigens produced in plants will be assayed with antibodies and tested in mice.


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