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Pubmed ID :29734743
Publication Date : //

Development of an Expression Vector to Overexpress or Downregulate Genes in .


, an endophytic fungus in the Ascomycota, provides plants with thermotolerance only when it carries a mycovirus known as thermotolerance virus (CThTV), and forms a three-way symbiotic relationship among these organisms. Under heat stress, several genes are expressed differently between virus-free (VF) and carrying CThTV (AN). We developed an expression vector, pM2Z-fun, carrying a zeocin resistance gene driven by the promoter, to study gene functions in to better understand this three-way symbiosis. Using this new 3.7-kb vector, five genes that are differentially expressed in —including genes involved in the trehalose, melanin, and catalase biosynthesis pathways—were successfully overexpressed or downregulated in VF or AN strains, respectively. The VF overexpression lines showed higher metabolite and enzyme activity than in the control VF strain. Furthermore, downregulation of expression of the same genes in the AN strain resulted in lower metabolite and enzyme activity than in the control AN strain. The newly generated expression vector, pM2Z-fun, has been successfully used to express target genes in and will be useful in further functional expression studies in other Ascomycota fungi.

Authors : Liu Chengke , Cleckler Blake , Morsy Mustafa ,

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