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

Salmonella typhi gene deletion strain, vaccine prepared from salmonella typhi gene deletion strain and application

A technology of Salmonella typhi and gene deletion strains, which is applied in the field of animal bacterial genetic engineering, can solve the problem that biosafety requirements cannot be used as vaccine strains, etc., and achieves the effects of broad market application prospects, clear genetic background and weak virulence

Active Publication Date: 2012-09-19
HENAN UNIV OF SCI & TECH
View PDF2 Cites 3 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, the use of transposon-mediated methods has randomness, and is only suitable for the screening of subtypes with significant phenotypic changes, and strains that do not cause significant phenotypic changes cannot be obtained through these methods; the traditional use of homologous recombination leads to Although the target gene method is accurate, the obtained strains often contain resistance markers in the end, and cannot be used as vaccine strains for vaccine production because they do not meet the biosafety requirements

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
  • Salmonella typhi gene deletion strain, vaccine prepared from salmonella typhi gene deletion strain and application
  • Salmonella typhi gene deletion strain, vaccine prepared from salmonella typhi gene deletion strain and application
  • Salmonella typhi gene deletion strain, vaccine prepared from salmonella typhi gene deletion strain and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The construction of embodiment 1 Salmonella typhi ST-1crp gene deletion strain

[0031] 1. Primer design (for gene cloning and molecular detection)

[0032] Reference literature (Yu Chuan, et al. Construction of Δcrp-deficient strain of Salmonella choleraesuis C78-1 strain and preliminary study on its biological characteristics. Journal of Animal Husbandry and Veterinary Medicine, 2010, 41(5): 587-593) and reported Salmonella typhi strains The crp gene sequence of Ty2 (GenBank No: AE016847) designed 2 pairs of primers (pr1 / pr2 and pr3 / pr4, see Table 1), from the virulent strain of Salmonella typhi ST-1 (ST-1 is the wild type typhi isolated from pigs) The virulent strain of Salmonella was preserved in the China Center for Type Culture Collection (CCTCC) on November 3, 2011, and the preservation number is: CCTCC NO: M2011372.) In the genome, the upstream and downstream fragments of crp gene crp1 (upper arm) and crp2 ( lower arm), the sizes of amplified fragments are 1048...

Embodiment 2

[0044] Example 2 Identification and biological characteristics of Salmonella typhi ST-1crp gene deletion strain ST-1

[0045] 1. Phenotype identification of gene deletion strain ΔcrpST-1

[0046] Inoculate the gene deletion strain ΔcrpST-1 and the parental strain ST-1 on a solid LB plate by streaking, and then transfer to glucose, maltose, lactose, sucrose, rhamnose, mannose, arabinose, xylose, dulcitol, urea, etc. Carbon source and H 2 Biochemical identification tubes such as S (purchased from Hangzhou Tianhe Company) were used for biochemical reactions. O and H antigens were identified according to the instructions of serum factors (purchased from China Veterinary Drug Administration). The results showed that the biochemical characteristics of the gene deletion strain ΔcrpST-1 were not completely consistent with the parent strain ST-1. Parental strain ST-1 can utilize maltose, and red colonies appear on MacConkey agar with maltose (see Figure 5 C), and after the crp gen...

Embodiment 3

[0051] Example 3 Salmonella typhi gene deletion vaccine

[0052] The obtained Salmonella typhi gene deletion strain ΔcrpST-1 was identified, each generation was inoculated on MacConkey agar medium containing 1% maltose to observe the colony color, and the crp gene of Salmonella typhi was used for PCR detection to identify the genetic stability of the deletion bacteria . After 50 passages, it was found that the colonies of the deletion strain ΔcrpST-1 could not turn red on the MacConkey agar medium containing 1% maltose, which still met the phenotypic characteristics of crp gene deletion; Stablize. The Salmonella typhi gene deletion strain ΔcrpST-1 was cultured on LB solid medium, and a single colony was picked and cultured in LB liquid medium until the concentration of viable bacteria reached 3.0×10 9 CFU / mL. Add gelatin protectant according to the ratio of bacteria liquid: gelatin protectant (volume: volume) (volume: volume) (this gelatin protectant preparation method is: ...

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 a Salmonella typhi gene deletion strain, a vaccine prepared from the Salmonella typhi gene deletion strain and application of the strain. The bacterial strain is Salmonella typhi delta crpST-1 and preserved in the China center for type culture collection, and the preservation number is CCTCC NO: M2011373. The Salmonella typhi gene deletion strain disclosed by the invention has excellent immunizing protection efficacy and weak toxicity without any adverse side effect on immunized pigs and is safer compared with the traditional vaccine. Therefore, the vaccine which is prepared from the gene deletion strain built by a parent strain has wide market application prospect. In addition, the Salmonella typhi gene deletion strain disclosed by the invention retains the immune protective efficacy aiming to salmonella typhi infection, and has clear genetic background and stable characteristics, so that the Salmonella typhi gene deletion strain has better biosecurity. The Salmonella typhi gene deletion strain disclosed by the invention does not contain a resistance tag and totally accords with the biological security requirement of vaccine.

Description

technical field [0001] The invention relates to a gene-deleted strain of Salmonella typhi without a resistance marker, a vaccine constructed by it and the application of the vaccine, which belong to the technical field of animal bacterial genetic engineering. Background technique [0002] Salmonella (Salmonella) is a Gram-negative bacillus parasitic in humans and animals. Salmonella is often isolated from humans, livestock, poultry, wild animals, and rodents, and bacteria of this genus also often exist in water, milk, meat, eggs, and other foods and feeds. Salmonellosis is a general term for different forms of humans, domestic animals and wild animals caused by various types of Salmonella. Some Salmonella are important zoonotic pathogens, some are animal disease pathogens, and some are common pathogenic bacteria for human and animal food poisoning or feed poisoning, so this genus of bacteria is very important in food hygiene, foreign trade export, and human and animal disea...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A61K39/112A61P31/04C12R1/42
Inventor 赵战勤薛云王臣王浩周金龙
Owner HENAN UNIV OF SCI & TECH
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