Čeština
EnglishTransient expression of heterologous model gene in plants using Potato virusX-based vector
Čeřovská, Noemi; Pečenková, Tamara; Moravec, Tomáš; Velemínský, Jiří
PLANT CELL, TISSUE AND ORGAN CULTURE 79 [2]: 147-152, 2004
Klíčová slova: plant virus; based vector; transient expression
Abstrakt: To optimize the efficiency of expression of foreign proteins using Potato virus X (PVX) -- based vector, the gene for the coat protein (CP) of other virus (Potato virusA, PVA) was cloned into the vector, propagated in E. coli and subsequently inoculated or agroinfected into the host plants. Host range studies showed that the best host plant is N. benthamiana. By means of RT PCR the presence and the stability of the construct were tested. Both ELISA and Western blot analysis were applicable for expressed protein detection. Expression level of PVA CP achieved approximately 5--10 per mille of total soluble proteins. The results demonstrated that agroinfection is the most suitable method for the propagation of our model gene using PVX--based vectors.
DOI:
Autoři z ÚEB: Noemi Čeřovská, Tomáš Moravec
PLANT CELL, TISSUE AND ORGAN CULTURE 79 [2]: 147-152, 2004
Klíčová slova: plant virus; based vector; transient expression
Abstrakt: To optimize the efficiency of expression of foreign proteins using Potato virus X (PVX) -- based vector, the gene for the coat protein (CP) of other virus (Potato virusA, PVA) was cloned into the vector, propagated in E. coli and subsequently inoculated or agroinfected into the host plants. Host range studies showed that the best host plant is N. benthamiana. By means of RT PCR the presence and the stability of the construct were tested. Both ELISA and Western blot analysis were applicable for expressed protein detection. Expression level of PVA CP achieved approximately 5--10 per mille of total soluble proteins. The results demonstrated that agroinfection is the most suitable method for the propagation of our model gene using PVX--based vectors.
DOI:
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