Novel gene engineering subunit vaccine for mycoplasma gallisepticum

A technology of mycoplasma gallisepticum and amino acids, which is applied in the direction of vaccines, veterinary vaccines, bacterial antigen components, etc., can solve the problems of lack, weak immune protection of vaccines, and easy mutation of antigenic proteins, so as to achieve sufficient supply and protein immunogen Good sex and good immunogenicity

Active Publication Date: 2019-07-12
苏州沃美生物有限公司
View PDF2 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, during the culture process of Mycoplasma gallisepticum, due to changes in conditions such as culture temperature, medium composition and self-replication, the antigenic protein on the surface of Mycoplasma gallisepticum is prone to mutation and loss, resulting in weak immune protection of inactivated vaccines The problem

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
  • Novel gene engineering subunit vaccine for mycoplasma gallisepticum
  • Novel gene engineering subunit vaccine for mycoplasma gallisepticum
  • Novel gene engineering subunit vaccine for mycoplasma gallisepticum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0143] Example 1 Construction and Identification of Transfer Vector pF-MGC1

[0144] 1. Amplification and purification of the MGC1 gene The codon-optimized MGC1 gene (SEQ ID NO: 1) was synthesized in Nanjing GenScript and cloned into the pUC17 vector to obtain the pUC-MGC1 plasmid vector. The pUC-MGC1 plasmid was used as a template, and MGC1-F and MGC1-R were used as upstream and downstream primers for PCR amplification (the gene sequences of MGC1-F and MGC1-R are shown in SEQ ID NO.11 and 12). For the amplification system, see Table 1.

[0145] Table 1 MGC1 gene amplification system

[0146]

[0147] The reaction conditions were: 95°C pre-denaturation for 5 minutes; 94°C denaturation for 45 seconds, 54°C annealing for 45 seconds, 72°C extension for 1 minute, 35 cycles; 72°C extension for 10 minutes.

[0148] Perform gel electrophoresis on the PCR product to verify the size of the target gene, such as figure 1 As shown, the target band appeared at the position of 2.9kbp,...

Embodiment 2

[0161] Example 2 Construction and Identification of Transfer Vector pF-MGC2

[0162] 1. Amplification and purification of the MGC2 gene The codon-optimized MGC2 gene (SEQ ID NO: 3) was synthesized in Nanjing GenScript and cloned into the pUC17 vector to obtain the pUC-MGC2 plasmid vector. The pUC-MGC2 plasmid was used as a template, and MGC2-F and MGC2-R were used as upstream and downstream primers for PCR amplification (the gene sequences of MGC2-F and MGC2-R are shown in SEQ ID NO.13 and 14). For the amplification system, see table 5.

[0163] Table 5 MGC2 gene amplification system

[0164]

[0165]

[0166] The reaction conditions were: 95°C pre-denaturation for 5 minutes; 94°C denaturation for 45 seconds, 54°C annealing for 45 seconds, 72°C extension for 1 minute, 35 cycles; 72°C extension for 10 minutes.

[0167] Perform gel electrophoresis on the PCR product to verify the size of the target gene, such as Figure 4 As shown, the target band appeared at the positi...

Embodiment 3

[0179] Example 3 Construction and Identification of Transfer Vector pF-MGC3

[0180] 1. Amplification and purification of the MGC3 gene The codon-optimized MGC3 gene (SEQ ID NO: 5) was synthesized in Nanjing GenScript and cloned into the pUC17 vector to obtain the pUC-MGC3 plasmid vector. The pUC-MGC3 plasmid was used as a template, and MGC3-F and MGC3-R were used as upstream and downstream primers for PCR amplification (the gene sequences of MGC3-F and MGC3-R are shown in SEQ ID NO.15 and 16). For the amplification system, see Table 9.

[0181] Table 9 MGC3 gene amplification system

[0182]

[0183] The reaction conditions were: 95°C pre-denaturation for 5 minutes; 94°C denaturation for 45 seconds, 54°C annealing for 45 seconds, 72°C extension for 1 minute, 35 cycles; 72°C extension for 10 minutes.

[0184] Perform gel electrophoresis on the PCR product to verify the size of the target gene, such as Figure 7 As shown, the target band appeared at the position of 2.7kbp...

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 an immunological composition and a subunit vaccine. The immunological composition comprises a protein which is selected from one or an arbitrary combination of two or more of mycoplasma gallisepticum-associated proteins encoded with nucleic acid molecules of SEQ ID NO: 1 or 3 or 5 or 7 or 9 or nucleic acid molecules which are 95% or above identical to the nucleotide sequenceof SEQ ID NO: 1 or 3 or 5 or 7 or 9. The vaccine adopts eukaryotic expression, the antigenicity and immunogenicity of the product are similar to those of a natural protein, the expression level is high, the immunogenicity is strong, the protective effect is good, and the vaccine has no pathogenicity to chickens; besides, large-scale serum-free suspension culture preparation of the vaccine can berealized through a bioreactor, and meanwhile the vaccine production cost is greatly reduced.

Description

technical field [0001] The application relates to the technical field of animal immunity drugs, in particular to a preparation method and application of a novel genetic engineering vaccine of Mycoplasma gallisepticum. Background technique [0002] Mycoplasma Gallisepticum (MG) infection is a kind of respiratory disease in chickens, which mainly causes chronic respiratory disease, air sacculitis and sinusitis. Clinically, it mainly manifests as cough, runny nose, rales when breathing, and mouth breathing in severe cases. According to statistics, after Mycoplasma gallisepticum infected chickens, the weak chick rate of chicks increased by about 10%, the laying rate of laying hens decreased by 10%-20%, the body weight of broilers decreased by 38%, the slaughter period was prolonged, and the feed conversion rate decreased by 21%. %, and can indirectly cause a large amount of medical expenses, it is one of the important diseases that endanger the chicken industry. [0003] Mycopl...

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): A61K39/02A61P31/04
CPCA61K39/0241A61P31/04A61K2039/552A61K2039/523
Inventor 曹文龙孔迪滕小锘易小萍张大鹤
Owner 苏州沃美生物有限公司
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