Construction method of gene engineering strain of Lactobacillus paracasei with high bacteriocin yield

The technology of a genetically engineered strain and a construction method is applied in the construction field of genetically engineered strains, and can solve the problems of low bacteriocin yield and titer of Lactobacillus paracasei.

Inactive Publication Date: 2016-07-13
HEILONGJIANG UNIV
View PDF4 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The present invention aims to solve the problem of low bacteriocin yield and titer of the existing Lactobacillus paracasei, and provides a method for constructing a genetically engineered strain of Lactobacillus paracasei with high bacteriocin production

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 method of gene engineering strain of Lactobacillus paracasei with high bacteriocin yield
  • Construction method of gene engineering strain of Lactobacillus paracasei with high bacteriocin yield
  • Construction method of gene engineering strain of Lactobacillus paracasei with high bacteriocin yield

Examples

Experimental program
Comparison scheme
Effect test

specific Embodiment approach 1

[0028] Specific embodiment one: the construction method of the Lactobacillus paracasei genetically engineered bacterial strain of high-yield bacteriocin of the present embodiment, carry out according to the following steps:

[0029] 1. Amplify the prcK gene from Lactobacillus paracasei HD1.7;

[0030] 2. Ligate the pMD18-T vector with the prcK gene to construct the plasmid pMD18-T-prcK;

[0031] 3. Using the expression plasmid pRSFDuet-1 as the backbone, insert the target gene prcK into the multiple cloning site on the plasmid to construct the recombinant overexpression vector pRSFDuet-1-prcK;

[0032] 4. Using the expression plasmid pRSFDuet-1-prcK as the backbone, insert the target gene tet into the multiple cloning site on the plasmid to construct the recombinant overexpression vector pRK-tet;

[0033] 5. The overexpression vector pRK-tet was electrotransformed into Lactobacillus paracasei HD1.7 competent cells;

[0034] 6. Screening and identification of prcK gene overex...

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 a construction method of a gene engineering strain of Lactobacillus paracasei with high bacteriocin yield, relates to a construction method of the gene engineering strain of the Lactobacillus paracasei and aims to solve the problem of low bacteriocin yield of the Lactobacillus paracasei at present. The method comprises steps as follows: step one, a prcK gene is amplified from Lactobacillus paracasei HD1.7; step two, a plasmid pMD18-T-prcK is constructed; step three, a recombinant overexpression vector pRSFDuet-1-prcK is constructed; step four, a recombinant overexpression vector pRR-tet is constructed; step five, the overexpression vector pRR-tet is used for performing electrotransformation on Lactobacillus paracasei HD1.7 competent cells; step six, overexpression mutant strains of the prcK gene are screened and identified. The bactericidal capacity of the gene engineering strain is increased by 17.86% compared with that of original strains; bacteriocin titer of fermentation broth after passage is 2372.46 plus or minus 46.27 AU / mL, and the titer after the passage is more stable; the method is applied to bacteriocin production.

Description

technical field [0001] The invention relates to a method for constructing a genetically engineered bacterial strain with high bacteriocin production. Background technique [0002] The production of Lactobacillus paracasei HD1.7 bacteriocin is closely related to the quorum effect, which is regulated by the three-component regulatory system (3CRS), including autoinducible peptide (AIP), histidine protein kinase ( HPK) and response regulator (RR). The product encoded by the prcK gene has homology with histidine protein kinase (HPK). Therefore, the overexpression of this gene will play a certain role in promoting the production of bacteriocins in Lactobacillus paracasei. However, there are few reports on the overexpression of L. paracasei gene, and there is still a blank in the use of overexpression technology to study the function of genes related to the quorum effect of Lactobacillus paracasei. Contents of the invention [0003] The invention aims to solve the problem of ...

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): C12N15/74C12N15/54C12N1/21C12R1/225
CPCC12N9/1205C12Y207/01037
Inventor 葛菁萍高冬妮孙艳阳王洋李兴霖纪晓磊杨睿睿平文祥
Owner HEILONGJIANG UNIV
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