Trichoderma harzianum acid A compound and trichoderma harzianum mutant strain producing same
A technology of Trichoderma harzianum and Trichoderma harzianum, which is applied in the field of fungal natural products, can solve the problems of poor experimental repeatability and inability to obtain genetic transformants, and achieve the effects of low cytotoxicity, high yield, and high anti-inflammatory activity
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[0038] As mentioned above, the existing research strategies for the discovery of fungal natural products are always unsatisfactory, and there are problems of one kind or another. For example, (1) the use of microorganisms in extreme environments requires heavy workload and low efficiency. A very small amount of new compounds; (2) heterologous expression requires the assembly of gene clusters, because the gene clusters are often large, and the experiment difficulty is high; (3) transcription regulation, chemical epigenetic modification is commonly used, and stable heritability cannot be obtained transformants, poor experimental repeatability. In order to find transformants with simple operation, good reproducibility, and stable heredity and high yield of functional natural compounds, the inventors conducted a lot of research on natural product mining strategies.
[0039] The present inventors have found that by knocking out the histone deacetylase gene HOS2 from Trichoderma har...
Embodiment 1
[0066] 1. Schematic diagram of constructing transformants by knocking out HOS2, such as Figure 9
[0067] 2. Construction of homology arm fragments
[0068] The homologous fragments of the target knockout gene were constructed using fusion PCR technology, the 5F and 3F fragments upstream and downstream of the target knockout gene were amplified according to the PCR system shown in Table 1, and analyzed by agarose gel electrophoresis and analyzed using the kit The fragments were recovered, and the recovered fragments were used as templates for fusion PCR, and homologous fragments were obtained by amplifying the PCR system as shown in Table 2. When the forward primer in Table 1 is 5F-F and the reverse primer is 5F-R, the upstream 5F fragment of the target knockout gene HOS2 is amplified; when the forward primer in Table 1 is 3F-F and the reverse primer is 3F- When R, the downstream 3F fragment of the target knockout gene HOS2 was amplified. The template DNA was the genomic D...
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