Date post: | 29-Jul-2015 |
Category: |
Science |
Upload: | marcinczyz |
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PROJECT
Plant-produced subviral HBV particles as carriers for antigen protein epitopes for the
purpose of vaccines with increased immunogenicity.
Worldwide need for a therapeutic vaccine against chronic HBV infections as THERE ARE OVER 370 MLN OF CHRONIC SICK worldwide who often develop liver dysfunction, cirrhosis and cancer.
Shepard CW et al. (2006)HBV infected population 2-8 %
< 2 %
> 8 %
Protein production in plants:
• Low cost of production – comparable with microbiological production
• Activity - proteins expressed in native and biologically active forms
• Safety – plants cannot transmit human pathogens
HBcAg Capsid-Like Particles
HBcAg as a carrier for immunogenic epitopes
Acc. James A. Perkins, 2002
• Plant–expressed HBcAg forms Capsid-Like Particles, very durable and immunogenic structures and a good platform for presenting immunogenic epitopes
• PreS2 domain - structural element of surface M- and L-HBs antigens, which induces high immunological response in 99,5% of population
PreS2 domain
Based on Wynne, 1999
HBcAg dimer
PreS2 domain
Research objectives
Transformation of Nicotiana tabacum to obtain genetically modified plants expressing HBcAg–preS2 protein
Analysis of HBcAg–preS2 expression level and form of obtained transformed plants.
Selection of highest expressing plants for material propagation and further tests.
Research progress
Preparation of vectors forAgrobacterium tumefaciens
Purification of antigens for injection
GMO of tobaccoproducing HBc-preS2 antigen
Generativeand/or vegetative
propagationPlant transformation
and regeneration
plazmid Ti
wektor binarny
T-DNA
chromosom
Plants analysis
0
100
200
300
4J/3 7H/712B/1013G/33
[mg/g FW of leaves]
HBcAg-preS2868pz
868pz
1400pz
1000pz750pz500pz
300pz
2000pz
1400pz
1000pz750pz500pz300pz
2000pz
3000pz4000pz6000pz
10000pz
Ścieżki:M – DNA size marker5 etc.
HBs – positive control
- trangenic plants L-N – non-transgenic plantsW – negative control
Transgenic plants PCR analysisHBcAg-preS2 content
in transgenic plants