A genetic manipulation strategy of Hansenula polymorpha and its application

A Hansenula polymorpha, a part of the technology, applied in the field of Hansenula polymorpha genetic manipulation strategy, can solve the problems of cell lethal effect, high false positive rate, low screening efficiency, etc., to achieve a wide range of applications and overcome low efficiency. Effect
CN103667274BActive Publication Date: 2016-05-04INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
Publication Date
2016-05-04

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Abstract

The invention discloses a hansenula polymorpha genetic operation strategy and application thereof. The hansenula polymorpha genetic operation strategy is characterized in that a DNA (Deoxyribonucleic Acid) fragment group used for knocking out a target gene of hansenula polymorpha consists of a DNA fragment A and a DNA fragment B; the DNA fragment A in the direction from end 5' to end 3' is structured as 5' end sequence of the target gene, m, mazF expression cassette, and part of resistance screening marker gene cassette fragment A, in sequence; the DNA fragment B in the direction from 5' end to 3' end is structured as part of resistance screening marker gene cassette fragment B, n, and 3' end sequence of the target gene, in sequence. According to the hansenula polymorpha genetic operation strategy, methanol induced toxic protein mazF has the effect of cutting mRNA (messenger Ribonucleic Acid), thus the purpose of knockout without a marker or a trace can be realized; the hansenula polymorpha genetic operation can be performed conveniently.
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Description

technical field

[0001] The invention relates to a genetic manipulation strategy of Hansenula polymorpha and its application. Background technique

[0002] Hansenula polymorpha (Hansenula polymorpha) is a kind of methanol-trophic yeast that can grow rapidly with methanol as the only carbon source and energy source. It has gene expression regulation and protein translation similar to higher eukaryotes like other yeasts In addition to the post-modification mechanism, it also has a unique mechanism that is beneficial to the high-efficiency expression of foreign proteins. For example, the endogenous methanol oxidase (methanoloxidase) gene promoter MOXp is a strong inducible promoter, which is not sensitive to the repression of glucose or glycerol. Under the condition of limitation or lack of glycerol, it can be induced by methanol to start the high-efficiency expression of exogenous genes, so cell culture and protein expression can be controlled by time, and the controllability o...

Claims

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