Expression of Recombinant gD2 Protein in Transgenic Tomato plants for development of a plant-derived vaccine against Human herpes virus 2
Expression of Recombinant gD2 Protein in Transgenic Tomato plants for development of a plant-derived vaccine against Human herpes virus 2
Aya El-Turkey1, A. K. El-Attar1, A. E. Aboulata1, B. Othman2 and K. A. El-dougdoug2
ABSTRACT
Human herpes virus 2 (HSV) is a DNA-viral genome and cause infection for both human and animal. HSV-1 and HSV-2 are causing infection to human. HSV-2 is a sexually-transmitted virus. HSV-2 is prevalence in European countries and also in Egyptian-closed societies like Upper Egypt. Egyptian isolate of HSV-2 was isolated and Glycoprotein D (HSV-2gD) subunit was amplified using specific PCR primers.PCR product was molecularly cloned in t E.coli cells using PCR2.1/TOPO/ TA cloning vector. HSV-2gD fragment was liberated from 1 μg recombinant plasmid by using the restriction endonuclease EcoRI. HSV-2gD was successfully sub-cloned into binary plant expression vector PBI-121. HSV-2gD subunit-containing binary vector PBI 121 were transformed into Agrobacterium tumifaciens LBA 4404 strain for Agro-inoculation. Tomato plants were transferred into regeneration medium and genetically transformed with the HSV-2gD-PB21 binary vector through Agrobacterium co cultivation. Agro-inoculated tomato plants were tested for the HSV-2gD insertion and transcription using both PCR and RT-PCR and the results were successful in getting insert-containing tomatoes against HSV-2gD. Specific expression of the introduced antigen construct at the RNA level was detected by RT-PCR, using specific primers for the HSV-gD2 transcript. Expression of the HSV-2 gD antigen was assayed by ELISA on protein extracts isolated from same transfected plant tissue samples as for the RT-PCR assays. Results showed that the HSV-gD2 construct was expressed at both the RNA and protein levels in the transgenic tomato plants. Finally, we report here, the use of stable transformation to develop transgenic tomatoes expressing the biologically active HSV-2 gD antigen (gD2). Thus, our results may have implications for the development of plant based vaccine production systems against HSV-2.
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