Expression vector of schizochytrium limacinum as well as construction method and application of expression vector

Abstract

The invention provides an expression vector of schizochytrium limacinum as well as a construction method and application of the expression vector, and belongs to the field of gene engineering. The expression vector comprises a pSc-18S rDNA (ribosomal deoxyribonucleic acid) vector, a combination of a promoter and a terminator of schizochytrium limacinum, and a gene sequence of a resistance-fluorescence double selection marker, the combination of the promoter and the terminator comprises at least one of the following combinations: a combination of a promoter PT and a terminator TT, a combination of a promoter PA1 and a terminator TA, a combination of a promoter PA2 and a terminator TA, and a combination of a promoter PE2 and a terminator TE. According to the present invention, with the efficient schizochytrium limacinum genome editing and transformant screening technology, the resistance-fluorescence double-selection marker is efficiently integrated to the schizochytrium limacinum chromosome through the gene gun in the fusion protein expression form, such that the gene editing element test and the transformant rapid screening can be performed based on the expressed fluorescence intensity; especially, when endogenous genes are expressed, the technical advantages are very prominent.

Description

technical field [0001] The present disclosure belongs to the field of genetic engineering, and particularly relates to an expression vector of Schizochytrium, a method for constructing the expression vector and its application. Background technique [0002] Schizochytrium belongs to the heterotrophic eukaryotes of the family Thraustochytrid, has the ability to rapidly divide and release zoospores, and can use a variety of carbon sources as substrates for high-density fermentation to produce a variety of unsaturated fatty acids , such as docosahexaenoic acid (docosatetraenoic acid, DHA). In recent years, it has been found that some Schizochytrium can also synthesize carotenoids, such as β-carotene and astaxanthin. Carotenoids can be used as antioxidants or colorants in the fields of food, health products, cosmetics and animal breeding, and have considerable application prospects. Since the synthesis mechanism and metabolic regulation mechanism of Schizochytrium astaxanthin ...

AI Technical Summary

Problems solved by technology

However, compared with the model strains, the genetic operating system of Schizochytrium is still in an immature stage, and there are still problems such as fewer gene editing elements and complex transformant screening, which restrict the gene editing efficiency of Schizochytrium and thus hinder Expanded application of Schizochytrium

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