Improved vectors for Agrobacterium mediated genetic manipulation of Hypholoma spp. and other homobasidiomycetes
| Autor: | Gary D. Foster, Suhad A.A. Al-Salihi, Andy M. Bailey, Thomas A. Scott |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Microbiology (medical) Agrobacterium Transgene Green Fluorescent Proteins 030106 microbiology GFP Microbiology 03 medical and health sciences Transformation Genetic Botany Promoter Regions Genetic Hypholoma promoter Molecular Biology Gene Genetics Expression vector biology fungi Hypholoma Agrobacterium tumefaciens biology.organism_classification Coculture Techniques Introns Transformation (genetics) 030104 developmental biology Genetic Engineering Agaricales Hypholoma fasciculare |
| Zdroj: | Al-Salihi, S A A, Scott, T A, Bailey, A M & Foster, G D 2017, ' Improved vectors for Agrobacterium mediated genetic manipulation of Hypholoma spp. and other homobasidiomycetes ', Journal of Microbiological Methods, vol. 142, pp. 4-9 . https://doi.org/10.1016/j.mimet.2017.08.014 |
| ISSN: | 0167-7012 |
| Popis: | The basidiomycete fungi Hypholoma fasciculare and H. sublateritium are both prolific producers of sesquiterpenes and triterpenes, some of which have relevant pharmaceutical properties. Although H. sublateritium has been transformed in the past, the low reported efficiencies highlighted the need for establishing an effective simple transformation system for these valuable species. We have optimized Agrobacterium tumefaciens-mediated transformation through testing various parameters in these two Hypholoma species, showing that a mixture of homogenized mycelia and Agrobacterium (strain LBA4404) co-cultivated for 84 h at 25 °C is optimal for efficient transformation in these basidiomycetes. This study also reveals the requirements for transgene expression, with the first report of GFP expression in these Hypholoma, the need for an intron for such transgene expression, and further demonstrates the functionality of the expression vector by its use in Clitopilus passeckerianus. This development of transformation system and expression constructs, can facilitate further genetic investigation such as gene functionality in these fungi. |
| Databáze: | OpenAIRE |
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