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.

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

CN114703201APending Publication Date: 2022-07-05LIFECOME BIOCHEM
3 Cites 0 Cited by

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • 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

Examples

Experimental program
Comparison scheme
Effect test

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...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
Login to View More

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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
05 Jul 2022
Publication
CN114703201A
IPC
C12N15/31; C07K14/195; C12N15/76; C12N1/21; C12R1/54
CPC
C07K14/195; C12N15/76
Inventors
陈守文; 金辉