Artificially-synthesized vitreoscilla hemoglobin gene, corresponding engineered strain and application

A technology of genetically engineered bacteria strains, Vitella hyaline, applied in the fields of bioengineering and genetic engineering, can solve problems such as the influence of recombinant bacteria that have not been investigated, and achieve the effect of increasing yield and increasing yield

Inactive Publication Date: 2019-08-09
枣庄市杰诺生物酶有限公司 +1
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  • Abstract
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AI Technical Summary

Problems solved by technology

Therefore, pIB139 can be introduced into Streptomyces by a simple and efficient method of conjugative transfer between genera, and can be integrated at the attB site of the Streptomyces chromosome through specific recombination, and replicate with the replication of the host chromosome. Zhao Guangrong et al. constructed the vgb gene in The downstream of PermE* in pIB139 was transformed into Streptomyces by protoplast method, which proved the feasibility of applying this method to Streptomyces, but did not investigate the effect of recombinant strain vgb expression on the synthesis of target metabolites

Method used

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  • Artificially-synthesized vitreoscilla hemoglobin gene, corresponding engineered strain and application
  • Artificially-synthesized vitreoscilla hemoglobin gene, corresponding engineered strain and application
  • Artificially-synthesized vitreoscilla hemoglobin gene, corresponding engineered strain and application

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Embodiment 1

[0036] (1) Codon modification of gene vgb and total synthesis of new gene vgbL

[0037] According to the usage frequency of codons in Streptomyces coelicolor genome, each codon in the natural sequence of the gene vgb was checked for the usage frequency, and the amino acid sequence was kept unchanged to generate a new gene vgbL. In order to facilitate the operation of subsequent experiments, Nde1 and Xba1 enzyme cutting sites were introduced at both ends of the gene vgbL, and 9 nucleotides were added to the 5' and 5' ends of the vgbL nucleotide sequence to form a restriction endonuclease Nde1 site And the protection base, 8 nucleotides are added at the 3' end of the vgbL nucleotide sequence to form the restriction endonuclease Xba1 site and the protection base. The whole gene of vgbL was synthesized, and the synthesized DNA was inserted into the corresponding site of the vector pIB139 vector, and the sequence proved to be correct. Among them, gene synthesis, vector insertion, ...

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Abstract

The invention relates to an artificially-synthesized vitreoscilla hemoglobin gene. The nucleotide sequence of the gene is shown in SEQ ID No.1, and the gene is marked as vgbL. The invention further provides a corresponding genetically-engineered strain for producing enramycin. The invention further provides a method for constructing the genetically-engineered strain for producing the enramycin. The artificially-synthesized vitreoscilla hemoglobin gene, the strain and the method have the advantages that the nucleotide sequence of the novel vitreoscilla hemoglobin gene vgbL which can be efficiently expressed in streptomyces is obtained, and the gene is artificially synthesized and transferred into fungicidal streptomyces ATCC21013; compared with an original wild strain, by means of the genetically-engineered strain, the yield of the enramycin is increased by 16.6%, and therefore, it can be seen that the vgbL can become an effective means of increasing the yield of antibiotics in variousstreptomyces.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to the field of genetic engineering, and specifically refers to an artificially synthesized hemoglobin gene of Vitiligo hyaline, corresponding engineering strains and applications. Background technique [0002] Streptomyces is an important genus of actinomycetes. It produces most of the known antibiotics and other biologically active substances. About 70% of antibiotics in nature are produced by Streptomyces and their close relatives. Enramycin (enramycin), also known as enramycin, anlemycin, and persistent mycin, is composed of 13 different types of 17 amino acid molecules. The amino acids form a cyclic polypeptide, and the fatty acid molecules are connected to the aspartic acid at the end. According to the difference of the terminal amino acid molecule, it is divided into enramycin A and enramycin B, and enramycin is a mixture of these two components. [0003] Enramycin is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12N15/76C07K14/805C12P21/02C12R1/465
CPCC07K7/08C07K14/805C12N15/76C12N2800/22
Inventor 许向阳魏东芝魏巍董凤英刘乐乐
Owner 枣庄市杰诺生物酶有限公司
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