Recombinant lactic acid bacteria strain expressing chicken infectious bursal virus vp2 protein and Salmonella outer membrane protein and application thereof
A technology of recombinant lactic acid bacteria and chicken infectivity, applied in the direction of virus/bacteriophage, virus, virus peptide, etc., can solve the problem of unsatisfactory effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0036] Construction and identification of embodiment 1 recombinant lactic acid bacteria
[0037] 1. Construction of recombinant lactic acid bacteria vector
[0038] The genes encoding chicken infectious bursal virus virus VP2 protein and Salmonella outer membrane protein (OMP) were optimized according to the codon preference table of lactic acid bacteria, and the optimized genes were named OptiVP2 and OptiOMP respectively. The optimized chicken Infectious bursal virus virus VP2 gene sequence OptiVP2 is shown in SEQ ID NO: 3, and the amino acid sequence of the encoded protein is shown in SEQ ID NO: 4, and the optimized outer membrane protein (OMP) of Salmonella gallinarum The gene sequence of OptiOMP is shown in SEQ ID NO:5, and the amino acid sequence of the encoded protein is shown in SEQ ID NO:6. The optimized OptiVP2 and OptiOMP gene sequences are connected through the link sequence (shown in SEQ ID NO.7) to obtain the nucleotide sequence encoding the VP2-OMP fusion protei...
Embodiment 2
[0049] Example 2 Detection of the adhesion characteristics of the fusion protein of VP2 protein expressed by recombinant lactic acid bacteria r-L.lactis-OMPH-VP2 of the present invention and OMP
[0050] Inoculate the recombinant lactic acid bacteria strain r-L.lactis-OMPH-VP2 in Elliker-medium medium, add 0.5% lactose, culture statically at 30°C overnight, subculture according to 1:50, and culture the bacteria density to OD 600 When it is 0.3-0.5, add nisin to induce culture, and the concentration of Nisin is 10ng / mL for culture. After static induction culture for 5 to 6 hours, the cell lysate supernatant is obtained after concentration and lysis, which contains the fusion protein of expressed OMP and VP2 protein .
[0051] At the same time, the recombinant lactic acid bacteria r-L.lactis-VP2 (the preparation method is the same as that of r-L.lactis-OMPH-VP2, which only expresses the VP2 protein of chicken infectious bursal virus virus) and the recombinant lactic acid bacteri...
Embodiment 3
[0053] The determination of the immunogenicity of embodiment 3 recombinant lactic acid bacteria of the present invention
[0054] Take out the recombinant Lactococcus lactis strain r-L.lactis-OMPH-VP2 prepared in Example 1 and the recombinant lactic acid bacteria r-L.lactis-VP2 that only express chicken infectious bursal virus virus VP2 protein (preparation method and The preparation of r-L.lactis-OMPH-VP2 is the same, the only difference is that it does not contain the OMP expression sequence), respectively streaked on the solid L-Elliker medium, cultured at 30°C, and after 10h-12h a single colony grows, use incinerated Bacteria were picked with a spear tip, inoculated in liquid L-Elliker medium, and cultivated to logarithmic phase; transferred to liquid L-Elliker medium at a ratio of 1:100 by volume and cultured at 30°C for 12h-14h , and then transferred to liquid L-Elliker medium at a ratio of 1:50 by volume and cultured at 30°C for about 2h-3h. When OD600=0.4-0.5, add Nisi...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
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