Prokaryotic expression method for sea cucumber cathepsin

A cathepsin and prokaryotic expression technology, which is applied in the field of prokaryotic expression of sea cucumber cathepsin, can solve the problems of sea cucumber cathepsin gene screening and functional research that have not yet been systematically carried out, and achieve the effect of easy control of the quality of expression products

Inactive Publication Date: 2015-08-19
ZHEJIANG OCEAN UNIV
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Problems solved by technology

However, the screening and functional research of sea cucumber cathepsin genes an...

Method used

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Embodiment

[0024] Embodiment: a kind of prokaryotic expression method of sea cucumber cathepsin, comprising, wherein: comprise to sea cucumber cathepsin gene sequence cloning; On the basis of obtaining sea cucumber cathepsin gene sequence clone, construct the fusion expression vector with sea cucumber cathepsin gene sequence, then The fusion expression vector is transformed into an expression host bacterium to induce the expression of the target protein; the sea cucumber cathepsin gene sequence contains the sequence of SEQ1. The expression host bacteria is Escherichia coli. The primers used to amplify the DNA fragments of the target protein used in the cloning of the sea cucumber cathepsin gene sequence are those that increase EcoRI and NotI recognition sites

[0025] Primer 1: F: 5'-CCGGAATTCATGCCAGACACTGTTGATT-3'; (SEQ3)

[0026] Primer 2: R: 5'-ATAGTTTAGCGGCCGCTTAGACAGTTGGGTAACTG-3'. (SEQ4)

[0027] The restriction endonucleases used to construct the fusion expression vector were E...

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Abstract

The invention discloses a prokaryotic expression method for sea cucumber cathepsin, the prokaryotic expression method comprises the following steps of carrying out cloning on gene sequence of the sea cucumber cathepsin; constructing fusion expression vector with the gene sequence of the sea cucumber cathepsin on the basis of obtaining the sea cucumber cathepsin cloned gene sequence; transforming the fusion expression vector to expression host strain for inducing the expression of target protein. The prokaryotic expression method provided by the invention has the advantages that escherichia coli is adopted to express the sea cucumber cathepsin at low cost, and the quality of expression product is easy to control.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a prokaryotic expression method of sea cucumber cathepsin. Background technique [0002] Procollagen is a structural protein of body tissue, and its many physiological functions have been systematically studied and widely recognized. With the rapid development of the collagen industry, the demand for protease, the key production factor in the collagen enzymatic hydrolysis process, is also increasing. At present, the commonly used enzymes generally have the disadvantage of low enzymatic hydrolysis efficiency of collagen, and the use of genetic engineering technology to develop new collagenase can make up for this fatal defect, and the quality of collagenase products produced by bioengineering technology is easy to control and the production cost is low. low. Apostichopus japonicus has a very strong autolysis ability, the essence of which is the efficient enzymolysis of proteases in ...

Claims

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Application Information

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IPC IPC(8): C12N9/64C12N15/70
CPCC12N9/6405
Inventor 王天明肖金星郑刚杨水兵余海霞
Owner ZHEJIANG OCEAN UNIV
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