Screening of Cucurbitaceae Family to Select Efficient Host Species for Transient Expression
Journal Title: International Journal of Advanced Biotechnology and Research. - Year 2016, Vol 7, Issue 1
Abstract
ABSTRACT The use of plant viral vectors for the transient expression of heterologous proteins offer a useful tool for the large-scale production of importance proteins, such as antibodies and vaccines. Virus vectors are beneficial because of the autonomous replication and high levels of gene expression in a short time. Selecting the most productive host, and expression system are the most important effective factors in production of recombinant proteins. The ZYMV virus specifically infects Cucurbitacea family. Therefore, in order to find the best host among different spices from Cucurbitacea family were studied. In this study, the hypothesis was the choice of plant host affects the foreign gene retention level by a Zuchiini yellow mosaic virus(ZYMV) vector expressing the green fluorescent protein (GFP).To accomplish this, a novel virus vector integrity bioassay was developed based on an old concept, whereby full-length cDNA of the ZYMV vector were rub-inoculated onto leaves of four species of Cucurbitacea family including Cucurbit, Cucumber, Melon and Cantaloupe and control plants (infected by ZYMV wild type). Survey expression levels of GFP in all infected plants were measured by several molecular and biochemical-based approaches. For investigates the GFP expression using fluorescence microscopy, RT-PCR, SDS-PAGE, Immunobloting and ELISA assays confirmed gfp gene was correctly transcribed and translated in the infected plants. This host dependent effect, suggests some plants may present a more suitable environment than the others to support or maintain optimal levels of virus vector-mediated foreign gene expression. Our results demonstrated, that the cucurbit spices had the highest expression level of GFP and it is possible to use ZYMV-based viral vectors for molecular farming and producing recombinant proteins in Cucurbitacea family.
Authors and Affiliations
Vahid Jalali Javaran1, 2, Alireza Shafeinia1*, Mokhtar Jalali Javaran2*, Esmaeil Ghasemi Gojani1 and Malihe Mirzaee2
Keywords
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