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Aspergillus nidulans strong promoter ANP1 and application thereof

A technology of Aspergillus nidulans and promoters, which is applied in the direction of DNA/RNA fragments, recombinant DNA technology, the use of vectors to introduce foreign genetic materials, etc., and can solve the problem of limited promoters

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, the promoters currently available for fungal secondary metabolite synthesis genes are very limited, so the isolation of promoters and the acquisition of strong and effective promoters play an important role in the development of more efficient heterologous expression systems

Method used

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  • Aspergillus nidulans strong promoter ANP1 and application thereof
  • Aspergillus nidulans strong promoter ANP1 and application thereof
  • Aspergillus nidulans strong promoter ANP1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1, ANP1 and ANP2 have promoter activity

[0048] 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.

[0049] 1. Preparation of target fragments

[0050] 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:

[0051] RE-WAUP-F: 5'-cagtctcgccgtcttgctc-3';

[0052] RE-wa-R: 5'-CCTCAACACCATATTTTTAATCCCATGTGgatcaggagaaggagagtcaagtcc-3'.

[0053] 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 an aspergillus nidulans strong promoter ANP1 and application thereof. The aspergillus nidulans strong promoter ANP1 disclosed by the invention is the following a) or b) or c): a) a DNA molecule shown in a sequence 1 in a sequence table, wherein the DNA molecule has a promoter function; b) a DNA fragment which has 75% or more identity with a nucleotide sequence defined by a) and has the promoter function; and c) a DNA fragment which is hybridized with the nucleotide sequence defined by a) or b) under strict conditions and has the promoter function. Experiments prove that the aspergillus nidulans strong promoter ANP1 disclosed by the invention has promoter activity, can drive the expression of a heterologous gene in aspergillus nidulans, and provides more choices for constructing a new heterologous expression system.

Description

technical field [0001] The invention relates to the strong promoter ANP1 of Aspergillus nidulans and its application 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 the acquisition of strong and effective promoters play an imp...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/81C12N15/11
CPCC07K14/38C12N15/81
Inventor 尹文兵余婧雯魏鹏霖
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI