Method for conversing aspergillus fumigatus and special-purpose expression carrier for the same

Abstract

The invention discloses a method used for transforming Aspergillus fumigatus and a special expression vector of Aspergillus fumigatus. The expression vector is a recombination expression plasmid prepared by inserting orotidine-5'-monophosphate decarboxylase gene to polyclone sites of Ti plasmid. The method used for transforming Aspergillus fumigatus of the invention is that: an exogenous DNA is inserted into the expression vector, so a recombination expression vector containing exogenous DNA is obtained, and is transformed into an Aspergillus fumigatus strain that has a function defective orotidine-5'-monophosphate decarboxylase gene through the mediation of agrobacterium tumefaciens. The method used for transforming Aspergillus fumigatus and a special expression vector of Aspergillus fumigatus of the invention are widely applied in removing the Aspergillus fumigatus, so as to study the fuction of desired genes.

Description

technical field [0001] The invention relates to a method for transforming Aspergillus fumigatus and a special expression carrier thereof. Background technique [0002] Invasive aspergillosis (Invasive aspergillosis, IA) IA is mainly caused by Aspergillus fumigatus infection. In recent years, the incidence of IA has shown an upward trend, and the condition is very serious. The mortality rate of IA in severe immunocompromised (immunocompromised) bodies such as leukemia and bone marrow transplant recipients often exceeds 90%. One of the main reasons. [0003] To clarify the pathogenesis of invasive aspergillosis, we must first understand the gene function of Aspergillus fumigatus, which determines the pathogenicity, pathogenesis, drug resistance and other traits of the fungus. The genome sequencing of Aspergillus fumigatus was completed at the end of 2005, which accelerated the research on gene function of Aspergillus fumigatus. [0004] Gene knockout is an important method ...

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Problems solved by technology

The disadvantages of using this selection marker are: hygromycin should be added to the medium for screening transformants; the transformation and screening processes are separate, and two types of media are required: basal medium and screening medium; Nylon membrane; In addition, in the experiment of Agrobacterium tumefaciens-mediated human pathogenic fungus—Histoplasma, it was found that some of the obtained transformants were pseudo-clones (Sullivan TD, Rooney PJ, Klein BS.Agrobacterium tumefaciens integrates transfer DNA into single chromosomal sites of dimorphic fungi and yields homokaryotic progeny from multinucleate yeast. Eukaryot Cell. 2002, 1(6): 895-905.), the reason is that some Aspergillus fumigatus will Induced drug resistance, the formation of pseudo-clones

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