Vitreoscilla hemoglobin expression cassette suitable for streptomyces fradiae and application thereof

A technology for Streptomyces flexneri, Vibrio vibrio, in applications, microbial-based methods, bacteria, etc.

Pending Publication Date: 2022-07-05
LIFECOME BIOCHEM
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  • Abstract
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  • Claims
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Problems solved by technology

At present, it is not possible to select a suitable signal peptide based on the previous results to construct the Vitreum hyaline hemoglobin expression cassette suitable for Streptomyces freundii for the high-efficiency expression of Vitreus hyaline hemoglobin in Streptomyces freundii and the metabolism based on the high-efficiency expression of Vitreum hyaline hemoglobin Efficient Synthesis of Products

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  • Vitreoscilla hemoglobin expression cassette suitable for streptomyces fradiae and application thereof
  • Vitreoscilla hemoglobin expression cassette suitable for streptomyces fradiae and application thereof
  • Vitreoscilla hemoglobin expression cassette suitable for streptomyces fradiae and application thereof

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Embodiment Construction

[0017] The specific embodiments of the present invention are described in detail below:

[0018] The technical solutions of the present invention, unless otherwise specified, are conventional solutions in the art; the reagents or materials, unless otherwise specified, are all derived from commercial channels.

[0019] Construction of Vibrio vibrio hemoglobin expression cassettes containing different Streptomyces signal peptides:

[0020] 1. vgbS is optimized according to the codon preference of Streptomyces flexneri, synthesized by Sangon Bio, and amplified by PCR to obtain the vgbS fragment; kasO* promoter, SP CynT Signal peptide, vgbS gene and T aph The four segments of the terminator are connected to form P kasO* -SP CynT -vgbS-T aph Expression cassette, its sequence is shown in SEQ ID NO.1.

[0021] 2. Using the pSET152 plasmid as a template, the pSET152 vector was obtained by PCR amplification, and the primers were G-T5-F and G-T5-R,

[0022]G-T5-F: atcgaattcgtaatc...

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Abstract

kasO*The invention belongs to the field of biotechnology and fermentation engineering, and provides a vitreoscilla hemoglobin expression cassette suitable for streptomyces fradiae and application thereof.The vitreoscilla hemoglobin expression cassette is obtained by connecting a vitreoscilla hemoglobin gene vgbS with a signal peptide SPCynT of carbonic anhydrase CynT in streptomyces fradiae, a streptomyces constitutive strong promoter PkasO * and a streptomyces fradiae terminator Taph; a VHb protein expression cassette PkasO *-SPCynT-vgbS-Taph is formed, and the nucleotide sequence of the expression cassette is as shown in SEQ ID NO. 1. The VHb protein expression cassette PkasO *-SPCynT-vgbS-Taph disclosed by the invention has more efficient VHb expression capacity, and when the VHb protein expression cassette PkasO *-SPCynT-vgbS-Taph is applied to streptomyces fradiae, the VHb expression effect is better, the fermentation product yield of the streptomyces fradiae is higher, and the yield of neomycin is at least increased by 14.9%.

Description

technical field [0001] The invention belongs to the field of biotechnology and fermentation engineering, and in particular relates to a oscillating bacillus hemoglobin expression cassette suitable for Streptomyces flexneri and its application. Background technique [0002] Streptomyces belong to the Streptomyces family of Actinomycetes of the prokaryotes kingdom taxonomically, with diverse genomes and extremely high GC (70-76%) content. About 70% of antibiotics in nature are produced by Streptomyces and its relatives Actinomycetes. Neomycin is an aminoglycoside antibiotic produced by Streptomyces flexneri, which can interfere with protein synthesis and is one of the most commonly used veterinary antibiotics at home and abroad. Its main components are neomycin B and C, and a small amount of neomycin A. Compared with other components, neomycin B has strong activity and low toxicity, and is the main component that needs to be monitored in the fermentation process. [0003] T...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C07K14/195C12N15/76C12N1/21C12R1/54
CPCC07K14/195C12N15/76
Inventor 陈守文金辉石姣魏子涵蔡冬波罗淦邱湘琪楼丽君李俊辉
Owner LIFECOME BIOCHEM
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