Construction of brucellosis A19 molecular marker vaccine strain and determination of virulence and immunogenicity
What is Al technical title?
Al technical title is built by PatSnap Al team. It summarizes the technical point description of the patent document.
A molecular marker vaccine, brucellosis technology, applied in the determination/examination of microorganisms, bacteria, recombinant DNA technology, etc. The effect of weak strength, improved vaccine function, and short market cycle
Active Publication Date: 2012-11-14
新疆维吾尔自治区畜牧科学院兽医研究所
View PDF2 Cites 9 Cited by
Summary
Abstract
Description
Claims
Application Information
AI Technical Summary
This helps you quickly interpret patents by identifying the three key elements:
Problems solved by technology
Method used
Benefits of technology
Problems solved by technology
In addition, the VirB12 gene is a virulence factor in the four-type secretion system of Brucella. The virulence of the A19 vaccine strain with VirB12 knockout has be
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
Click on the blue label to locate the original text in one second.
Reading with bidirectional positioning of images and text.
Smart Image
Examples
Experimental program
Comparison scheme
Effect test
Example Embodiment
[0034] Example 1
[0035] Construction of recombinant suicide plasmid
[0036] Materials and Methods
[0037] Strains, plasmids, vectors
[0038] Brucella A19 strain was purchased from the China Institute of Veterinary Drug Control, and the suicide plasmid pBK-CMV-sacB was preserved in the Ward Room of the Veterinary Research Institute of Xinjiang Academy of Animal Science; the cloned pGEM-T vector was a commercial reagent and was purchased from Promega.
[0039] Reagent
[0040] Restriction endonucleases were purchased from Fermentas; Plasmid DNA extraction kits and DNA gel recovery kits were purchased from Promega; T4DNA ligase, DNA Marker, and DH5α were products of Beijing Zhuangmeng Biotech Co., Ltd.; sequencing was determined by Shanghai Invitrogen Trading Limited completion.
[0041] Design primers according to GenBankAF141604 sequence, as shown in Table 1:
[0042] Table 1 Primer sequence
[0043]
[0044] PCR amplification
[0045] Amplification of the upstream homology arm segment ...
Example Embodiment
[0067] Example 2
[0068] Said is the determination of the virulence and immunogenicity of the Brucellosis A19 molecular marker vaccine strain.
[0069] Characteristics of A19 molecular marker strain:
[0070] The experimental animals were BALB / C female mice aged 4-6 weeks, purchased from Xinjiang Experimental Animal Research Center, license number: SCXK (new) 2003-0002.
[0071] Safety test: Dilute A19-ΔVirB12 live bacteria with normal saline to contain 1 billion live bacteria per 1ml, and subcutaneously inject 5 BABL / C mice weighing 18-20g, each with 0.25ml, and all are healthy within 10 days.
[0072] Genetic stability:
[0073] Passaging stability: Streak culture of the frozen A19-ΔVirB12 molecular marker strain on tryptone soy broth plate at 37℃ with 5% CO 2 After 72 hours of culture in an incubator, the colonies grown are first-class seeds. The first-class seeds are passed on the medium for 50 consecutive generations. The colonies of the passaged bacteria are selected every 5 gener...
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
Login to view more
Abstract
The invention relates to construction of a brucellosis A19 molecular marker vaccine strain and determination of virulence and immunogenicity. According to the invention, a suicide-plasmid-mediated homologous recombination technology is utilized to knock out a virulence factor VirB12 gene of a brucellosis tetratype secretory system of a brucellosis A19 vaccine strain, thus obtaining the A19-delta VirB12 molecular marker vaccine strain. Animal experiments show that the virulence of the A19 molecular marker vaccine strain is slightly weaker than that of a parent A19 vaccine strain, so that the strain safety is further improved, but the good immune effect on brucellosis is maintained; and moreover, the molecular marker vaccine strain has stable heredity. According to the invention, the brucellosis VirB12 gene is taken as a molecular marker, a polymerase chain reaction (PCR) method is applied to identify the brucellosis vaccine strain from wild viral strains, so that the molecular marker is provided for building a determination method for identifying vaccine immunization and natural infection, and the function of the original brucellosis A19 vaccine is improved, therefore, the brucellosis A19 molecular marker vaccine strain has an very important practical meaning on controlling the epidemic brucellosis, and has wide application prospect.
Description
technical field [0001] The invention relates to the construction of a brucellosis A19 molecular marker vaccine strain and the determination of virulence and immunogenicity, and belongs to the technical field of microbial bacterial genetic engineering. Background technique [0002] Brucellosis (brucellosis, referred to as brucellosis) is a zoonotic infectious disease mainly infecting domestic animals caused by Brucella. Brucella has the characteristics of wide range of hosts, strong infectivity and difficult cure after infection, which poses a serious threat to animal husbandry and human health. [0003] Vaccine immunization is the main measure to prevent and control brucellosis. At present, the A19 vaccine used to immunize cattle against brucellosis has relatively good immunity. This vaccine has been used in my country for more than 60 years, providing an important guarantee for the effective control of brucellosis, but it also has certain defects. . That is, the current A...
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
Application Date:The date an application was filed.
Publication Date:The date a patent or application was officially published.
First Publication Date:The earliest publication date of a patent with the same application number.
Issue Date:Publication date of the patent grant document.
PCT Entry Date:The Entry date of PCT National Phase.
Estimated Expiry Date:The statutory expiry date of a patent right according to the Patent Law, and it is the longest term of protection that the patent right can achieve without the termination of the patent right due to other reasons(Term extension factor has been taken into account ).
Invalid Date:Actual expiry date is based on effective date or publication date of legal transaction data of invalid patent.
Login to view more 
Login to view more