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A codon-optimized α-amylase gene of Yuzu sea cucumber and its recombinant expression vector and application

A technology of codon optimization and yuzu sea cucumber, applied in the direction of carrier, application, glycosylase, etc., can solve the problem that the α-amylase gene cannot be expressed successfully, and achieve the effect of improving feeding and digestion and absorption efficiency

Active Publication Date: 2021-05-25
SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the α-amylase gene of sea cucumber yuzu cannot be successfully expressed in the prokaryotic expression system, so genetic modification is needed to express the α-amylase gene of sea cucumber yuzu in the prokaryotic expression system

Method used

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  • A codon-optimized α-amylase gene of Yuzu sea cucumber and its recombinant expression vector and application
  • A codon-optimized α-amylase gene of Yuzu sea cucumber and its recombinant expression vector and application
  • A codon-optimized α-amylase gene of Yuzu sea cucumber and its recombinant expression vector and application

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Embodiment 1

[0034] 1. Acquisition of full-length amylase gene of sea cucumber Yuzu and prediction of open reading frame

[0035] The partial sequence of the α-amylase gene was screened from the second-generation transcriptome database of Yuzu sea cucumber constructed by our research group, and the full-length sequence of the gene was obtained by RACE technology. Then in the NCBI database ( https: / / www.ncbi.nlm.nih.gov / orffinder / ) to predict its open reading frame (ORF). The results showed that the full-length cDNA of the amylase gene was 1734bp, the open reading frame (ORF) was 1578bp, encoded 525 amino acids, and the predicted molecular weight was 59.34KDa. The protein contains a signal peptide and an amylase functional domain at the N-terminus, including the catalytic active site ( figure 1 ).

[0036] 2. Codon optimization of α-amylase gene of sea cucumber Yuzu and construction of protein expression vector

[0037] 2.1 Optimization of codons of α-amylase gene in sea cucumber Y...

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Abstract

The invention provides a codon-optimized α-amylase gene of sea cucumber yuzu and its recombinant expression vector and application. On the basis of the α-amylase gene sequence of sea cucumber yuzu, the codons of the first 20 amino acids after the signal peptide of the gene are optimized, and the nucleotide sequence of the optimized α-amylase gene of sea cucumber yuzu is shown as SEQ ID NO .1 shown. The amino acid sequence encoded by the optimized yuzu sea cucumber amylase gene sequence of the present invention is consistent with the original gene sequence, but the protein with α-amylase activity can be successfully expressed in the prokaryotic expression system (Escherichia coli BL21). Unoptimized α-amylase sequences cannot be expressed in prokaryotic expression systems.

Description

technical field [0001] The invention relates to the technical field of genetic modification and protein expression, in particular to a codon-optimized α-amylase gene of sea cucumber yuzu and its recombinant expression vector and application. Background technique [0002] Amylase is a hydrolytic enzyme, which is the most widely used enzyme in the fermentation industry. Amylase generally acts on α-1,4-glucans such as soluble starch, amylose and glycogen, and hydrolyzes their α-1,4-glycosidic bonds. According to the mode of action, amylases can be divided into alpha-amylases and beta-amylases. α-amylase can hydrolyze the α-1,4-glucosidic bond in starch, cut the starch into short-chain dextrins of different lengths and a small amount of low-molecular sugars, so that the viscosity of starch paste decreases rapidly and plays a role Reduces consistency and "liquefies" the action, thereby improving the ability of the animal's intestines to digest food and absorb nutrients. In fis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/56C12N15/11C12N15/70C12N1/21C12N9/26C12R1/19
CPCC12N9/2414C12N15/70C12N2800/22C12Y302/01001
Inventor 任春华吴小芬阮尧陈廷江晓
Owner SOUTH CHINA SEA INST OF OCEANOLOGY - CHINESE ACAD OF SCI