Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Antibacterial recombination polypeptide and expression vector thereof

A technology for recombinant polypeptides and vectors, applied in the field of genetic engineering, can solve problems such as unsuitable for large-scale production, cumbersome production process, and high cost

Inactive Publication Date: 2013-06-12
苗青青
View PDF2 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] In order to solve the cumbersome and expensive production process of Lactoferampin (Lfcin) and a chimeric structure peptide chain (N-FKCRRWQWRMKKLG-K-RSKNKGFKEQAKSLLKWILD-N) of Lactoferampin (Lfcin) and Lactoferampin (Lfampin) synthesized by artificial protein peptides , the yield is low, and it is not suitable for large-scale production. The present invention provides an antibacterial recombinant polypeptide and its expression vector

Method used

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
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Antibacterial recombination polypeptide and expression vector thereof
  • Antibacterial recombination polypeptide and expression vector thereof
  • Antibacterial recombination polypeptide and expression vector thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0081] The acquisition and identification of embodiment 1 recombinant plasmid pGE-Lfa / Lfc

[0082] Pick a single clone from the puncture bacteria, inoculate it into 5ml LB liquid medium containing 100ug / ml Amp, at 37°C, 220r / min, shake culture for 12-16h, OD value ≈ 0.6; small dose according to SDS alkaline lysis method Extract the recombinant plasmid pGE-Lfa / Lfc containing the antibacterial recombinant polypeptide Lfa-AA-Lfc gene fragment, use restriction endonuclease Hind Ⅲ to digest the recombinant plasmid pGE-Lfa / Lfc, and use 1.2% agarose gel electrophoresis to detect .

Embodiment 2

[0083] Example 2 Connection and Transformation of the Gene of the Antibacterial Recombinant Polypeptide Lfa-AA-Lfc and the Secreted Expression Vector pPIC9K

[0084] The gene sequence of the synthetic antibacterial recombinant polypeptide Lfa-AA-Lfc was used as a template, and P1 and P2 were used as upstream and downstream primers for PCR amplification. Anneal for 30s, extend for 30s at 72°C, 30 cycles, and finally extend for 3min at 72°C to obtain PCR amplification products; PCR amplification products were identified by agarose gel electrophoresis, ligated with pGEM-T Easy Vector, and transformed into Escherichia coli C600 Competent cells; Recombinant plasmid TE-Lfa-AA-Lfc was digested with restriction endonuclease EcoR Ⅰ, the antibacterial recombinant polypeptide Lfa-AA-Lfc gene fragment was recovered, filled in with Klenow Fragment enzyme; secreted by Pichia pastoris The type expression vector pPIC9K was recovered by restriction endonuclease SnaB Ⅰ digestion, and the two re...

Embodiment 3

[0086] Example 3 Linearization of recombinant plasmid pPIC9K-Lfa-AA-Lfc

[0087] The recombinant plasmid pPIC9K-Lfa-AA-Lfc was digested with the restriction endonuclease Sal I. After the digestion reaction, the linearized recombinant plasmid pPIC9K-Lfa-AA-Lfc was recovered with a Biospin gel recovery kit, and 0.8 % agarose gel electrophoresis to estimate the concentration of recombinant plasmid pPIC9K-Lfa-AA-Lfc.

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

No PUM Login to View More

Abstract

The invention relates to antibacterial recombination polypeptide and an expression vector of the antibacterial recombination polypeptide. The problems of being trivial in production technology, high in cost, low in yield and not suitable for mass production in an existing method of artificially synthesizing protein peptide are solved. According to the existing method of artificially synthesizing protein peptide, by the adoption of the method, a chimeric-structure peptide chain (N-FKCRRWQWRMKKLG-K-RSKNKGFKEQAKSLLKWILD-N) of Lactoferampin (Lfcin) and Lactoferampin (Lfampin) is synthesized. The gene nucleotide sequence of the antibacterial recombination polypeptide is shown as SEQ ID NO: 1 in a sequence table, and the amino acid sequence of the antibacterial recombination polypeptide is shown as SEQ ID NO: 2 in the sequence table. The antibacterial recombination polypeptide and the expression vector of the antibacterial recombination polypeptide have the advantages of being good in antibacterial effect, simple in production technology, low in cost, high in yield and suitable for mass production.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to an antibacterial recombinant polypeptide and its expression vector. Background technique [0002] In 1928, British bacteriologist Fleming discovered penicillin, which opened the prelude to the use of antibiotics in humans. Since the use of antibiotics, although countless lives have been saved, antibiotics have certain toxic and side effects. Long-term non-standard use can easily cause bacteria and strains to develop drug resistance, and at the same time inhibit the immune function of the body. The World Health Organization recommends that antibiotics be used in hospitals. The utilization rate does not exceed 30%, while in China it is as high as 74%. [0003] In recent years, antibiotics have been widely used not only in disease prevention and control, but also in food, health care products and animal feed without restriction, resulting in a decline in the use of anti...

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
no application Login to View More
IPC IPC(8): C12N15/62C07K19/00C12N15/81C12N15/66C12R1/84
Inventor 苗青青
Owner 苗青青
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com