Construction of lactobacillus acidophilus S-layer protein surface display system

A surface display system, Lactobacillus acidophilus technology, applied in biochemical equipment and methods, material inspection products, applications, etc., can solve the problem of no protein expression and achieve the effect of weakening the degradation effect

Inactive Publication Date: 2010-02-03
CHANGCHUN UNIV OF SCI & TECH
View PDF0 Cites 15 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The slpC gene was isolated from the slpB gene. Sequence analysis showed that the slpC gene has a complete promoter and terminator, and has the ability to encode SlpC, and the slpC gene can be detected from the

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
  • Construction of lactobacillus acidophilus S-layer protein surface display system
  • Construction of lactobacillus acidophilus S-layer protein surface display system
  • Construction of lactobacillus acidophilus S-layer protein surface display system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] (1) Acquisition of the S-layer SlpA gene of Lactobacillus acidophilus

[0079] 1 Materials and reagents

[0080] (1) strain

[0081] Lactobacillus acidophilus strain (Lactobacillus acidophilus 1.1878), Escherichia coli (E. coli JM109).

[0082] (2) Enzymes and main biochemical reagents

[0083] pMD18-T Vector System, Restriction Enzyme Sac I, Kpn I, dNTP, Ex Taq DNApolymerase, RNaseA, λ-Hind III Digest Marker, DL2000 Marker, Lysozyme, DNA Gel Recovery and Purification Kit, IPTG, Agar sugar.

[0084] 2 PCR amplification of SlpA gene

[0085] After the revived Lactobacillus acidophilus strains were cultured overnight at 37°C constant temperature anaerobic culture, a single colony was picked and inoculated in fresh MRS liquid medium for cultivation. 600 When the value is 0.6-0.8, collect the bacteria, use the chromosome extraction kit, and extract the chromosome DNA according to the instructions. According to the sequence of the published S-layer gene, a pair of prim...

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 provides construction of a lactobacillus acidophilus S-layer protein surface display system, which belongs to the field of biotechnology. The construction is characterized in that gene acquisition comprises (1) PCR amplification, (2) the connection of a SlpA gene and a pMD18-TVector system and (3) the identification of cloning plasmid pMD18T-S, and the construction comprises (1) theacquisition of a target gene and a double-labeling expression vector, (2) the connection and transformation of the double-labeling expression vector and SlpA, (3) the identification of a surface display vector system and (4) the detection of the anchored expression situation of S-layer protein on recombinant surface. The construction has the advantages that: 1, the constructed vector can be used to be transformed into S-layer protein gene defect lactobacillus to study the formation mechanism of lactobacillus S-layer protein; and 2, by utilizing a DNA recombination technique, protective antigengenes of pathogens are fused with lactobacillus S-layer protein on the surface display vector.

Description

technical field [0001] The invention belongs to the field of biotechnology. Background technique [0002] 1 Microbial cell surface display technology [0003] Microbial cell-surface display (MCSD) refers to the use of techniques such as molecular biology and immunology to anchor and display exogenous protective peptide molecules and protein molecules on the surface of microbial cells. The former is fused with a certain surface protein of the cell, and the former is positioned on the cell surface by means of the surface recognition and surface positioning functions of the bacterial protein. The former is called the passenger protein, and the latter is called the carrier protein. The expressed gene product does not need to be extracted. , purification, renaturation and other operations, you can directly use the whole cell to carry out the follow-up work of expressing the protein [1] . [0004] 2 Research Status of Surface Display System [0005] 2.1 Phage surface display s...

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): C12N15/31C12N15/70G01N33/53G01N21/64
Inventor 王春凤李景梅刘琼田坚
Owner CHANGCHUN UNIV OF SCI & TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap