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A kind of promoter and its application of starting the expression of Phaffia rhodozyma RNA

A Phaffia rhodozyma and promoter technology is applied in the fields of yeast biotechnology and synthetic biology to achieve the effects of improved gene editing efficiency, strong initiation ability and high gRNA expression level

Active Publication Date: 2022-05-17
QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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Problems solved by technology

However, this technique has not been reported in economically important basidiomycete yeasts

Method used

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  • A kind of promoter and its application of starting the expression of Phaffia rhodozyma RNA
  • A kind of promoter and its application of starting the expression of Phaffia rhodozyma RNA
  • A kind of promoter and its application of starting the expression of Phaffia rhodozyma RNA

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Experimental program
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Embodiment

[0035] 1. Materials and Methods

[0036] 1.1 Strains and culture conditions

[0037] Phaffia rhodozyme strain CBS 6938 (purchased from the Dutch CBS Culture Collection) was used as the material. Routine culture uses YPD liquid medium (glucose 20g / L, peptone 20g / L, yeast extract 10g / L), and the culture conditions are 22°C and 250rpm.

[0038] 1.2 Prediction of RNA polymerase III promoter

[0039] Taking the promoter sequences of SCR1 and U6 genes of Saccharomyces cerevisiae and Candida albicans as reference, and using the Optimize for Somewhat similar sequences program in the NCBIBlastn program package, the genome data of Phaffia rhodozyme Xanthophyllomyces dendrorhous (taxid: 5421) were obtained. Homologous sequences of the Phaffia rhodozyme genome. Using the same method, using the Saccharomyces cerevisiae transfer RNA (tRNA) sequence as a reference, the homologous sequence of the Phaffia rhodozyma tRNA gene was obtained through NCBI Blastn sequence alignment; the obtained ...

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Abstract

The invention relates to the fields of yeast biotechnology and synthetic biology, in particular to a Phaffia rhodozyma RNA polymerase III promoter and application thereof. The promoter is the Phaffia rhodozyma RNA polymerase III promoter; the promoter is SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, or 70% of the above sequences Homologous DNA nucleotide sequence with promoter activity. The promoter disclosed in the present invention is applied to the Phaffia rhodozyma CIRSPR-Cas9 system to guide the expression of gRNA in the system, and the gene editing efficiency is increased by 4 times.

Description

technical field [0001] The invention relates to the fields of yeast biotechnology and synthetic biology, in particular to a Phaffia rhodozyma RNA polymerase III promoter and application thereof. Background technique [0002] Yeast is of great value to scientific research and human life. Since the beginning of the 21st century, research results using Saccharomyces cerevisiae as a model have won the Nobel Prize in Physiology or Medicine in 2001, 2013 and 2016 respectively. In terms of industry, according to BCC Research statistics, the global market for Saccharomyces cerevisiae and related products reached US$7.1 billion in 2016, and is expected to reach US$10.07 billion by 2020. In addition to Saccharomyces cerevisiae, there are a large number of yeasts in nature. These non-model yeasts have rich species diversity, genetic diversity and metabolic diversity, but their application development is still very weak. [0003] With the development of genome sequencing and modern bi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/81C12N1/19C12R1/645
CPCC07K14/39C12N15/815
Inventor 王士安张宁李福利
Owner QINGDAO INST OF BIOENERGY & BIOPROCESS TECH CHINESE ACADEMY OF SCI
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