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Promoter derived from aspergillus nidulans and application of promoter in driving expression of heterologous genes in aspergillus nidulans

A technology of Aspergillus nidulans and promoters, which is applied in the biological field and can solve problems such as limited promoter development

Inactive Publication Date: 2021-06-18
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the field of fungi, the development of promoters is still limited

Method used

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  • Promoter derived from aspergillus nidulans and application of promoter in driving expression of heterologous genes in aspergillus nidulans
  • Promoter derived from aspergillus nidulans and application of promoter in driving expression of heterologous genes in aspergillus nidulans
  • Promoter derived from aspergillus nidulans and application of promoter in driving expression of heterologous genes in aspergillus nidulans

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Experimental program
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Embodiment 1

[0048] Example 1, ANP1 and ANP2 have promoter activity

[0049] This example verifies the activities of the two strong promoters ANP1 and ANP2 screened in the example, and the sequences of ANP1 and ANP2 are sequences 1 and 2 in the sequence listing, respectively.

[0050] 1. Preparation of target fragments

[0051] PCR amplification was carried out using the genomic DNA of Aspergillus nidulans LO4389 as a template, and the obtained PCR product was recorded as PCR product 1, and the primers used were as follows:

[0052] RE-WAUP-F: 5'-cagtctcgccgtcttgctc-3';

[0053] RE-wa-R: 5'-CCTCAACACCATATTTTTAATCCCATGTGgatcaggagaaggagagtcaagtcc-3'.

[0054] Taking pYWB2 (Zhang, P., Wang, X., Fan, A., Zheng, Y., Liu, X., Wang, S., Zou, H., Oakley, B.R., Keller, N.P., and Yin, W.B. (2017). A cryptic pigment biosynthetic pathway uncovered by heterologous expression is essential for conidial development in Pestalotiopsis fici. Mol Microbiol 105,469–483.) was used as a template for PCR ampli...

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Abstract

The invention discloses a promoter derived from aspergillus nidulans and application of the promoter in driving expression of heterologous genes in the aspergillus nidulans. The promoter (ANP2) derived from the aspergillus nidulans is the following (a), (b) or (c) that (a), a DNA molecule is as shown in a sequence 2 in a sequence table, and the DNA molecule has a promoter function; (b), a nucleotide sequence limited by the (a) has 75% or more than 75% of identity and has a DNA fragment of the promoter function; and (c), the nucleotide sequence limited by (a) or (b) is hybridized under strict conditions, and a DNA fragment of the promoter function is achieved. Experiments prove that the aspergillus nidulans strong promoter ANP2 has promoter activity, can drive the expression of the heterologous genes in aspergillus nidulans, and provides more choices for constructing a new heterologous expression system.

Description

technical field [0001] The invention relates to a promoter derived from Aspergillus nidulans and its application in driving heterologous gene expression in Aspergillus nidulans in the field of biotechnology. Background technique [0002] The abundant secondary metabolites produced by fungi are one of the important sources of modern natural medicines. Genome sequencing and bioinformatics predictions show that there are a large number of gene clusters for the biosynthesis of secondary metabolites in fungi, and most of them are silent. How to effectively activate these silent gene clusters is a scientific issue that researchers are concerned about. Promoters, as basic transcriptional regulatory elements, are widely used in the construction of high-efficiency heterologous expression systems and the transformation of synthetic pathways. However, the promoters currently available for fungal secondary metabolite synthesis genes are very limited, so the isolation of promoters and t...

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Application Information

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IPC IPC(8): C12N15/113C12N15/80C12R1/66
CPCC07K14/38C12N15/80
Inventor 尹文兵马紫卉余婧雯
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI