Maziar Habibi-Pirkoohi; Afsaneh Mohkami
Abstract
Green plants have emerged as ideal platforms for production of recombinant vaccine during recent decades. Various antigens relating to a large number of animal and human diseases have been studied in different plant species for production of recombinant vaccines. Despite the unique advantages of plant ...
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Green plants have emerged as ideal platforms for production of recombinant vaccine during recent decades. Various antigens relating to a large number of animal and human diseases have been studied in different plant species for production of recombinant vaccines. Despite the unique advantages of plant systems as green factories for production of recombinant vaccines, there are some major hurdles that have prevented commercial production of plant-based vaccines. In this review, theoretical background and practical applications of plant system for production of various recombinant vaccines are discussed.
Jalal Soltani; Mahsa Yousefi-Pour H.; Sonbol Nazeri
Abstract
The fungus Colletotrichum gloeosporioides is the causative agent of anthracnose disease of many tropical, subtropical and temperate fruits, and a microbial source of the anticancer drug, Taxol. Here, we introduce an optimized Agrobacterium tumefaciens-mediated transformation (ATMT) protocol for genetic ...
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The fungus Colletotrichum gloeosporioides is the causative agent of anthracnose disease of many tropical, subtropical and temperate fruits, and a microbial source of the anticancer drug, Taxol. Here, we introduce an optimized Agrobacterium tumefaciens-mediated transformation (ATMT) protocol for genetic manipulation of this fungus using hph and gfp-tagged hph genes as selection markers. Results showed that falcate spores can be easily used instead of protoplasts for transformation. Several experimental parameters were shown to affect transformation efficiencies, among which the length of co-cultivation, the ratio of fungal conidia to bacterium during co-cultivation, the kind of membrane during co-cultivation, and the kind of fungal growth medium during transformants selection, showed the highest influences on ATMT frequencies. Results indicated that the optimal ATMT of C. gloeosporioides was achived after 3 days of co-cultivation, at 107 per ml fungal conidia, via the use of Fabriano 808 filter paper and Czapek's culture medium. Successive subculturing of transformants on selective and non-selective media demonstrated the stable expression of transgens, and subsequent PCR based analyses of transformants revealed the presence (100%) of transferred genes. Flourescence microscopy analyses showed a punctuate pattern for localization of an expressed Gfp-tagged Hph protein inside fungal hyphae. The optimized ATMT protocol generated mutants that showed different phenotypes based on their vegetation and pigmentation. This suggests the possible applicability of this technique for functional genetics studies in C. gloeosporioides, through insertional mutagenesis.
