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Self-polymerized peptide fused CD151 protein as well as preparation method and application thereof

A technology of fusion protein and protein, applied in the field of biotechnology research

Inactive Publication Date: 2016-08-31
江苏泰州动物药品工程技术中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although this strategy is easy to think of, it faces the following technical problems in the specific design: whether the self-polypeptide fusion CD151 protein can be expressed in Escherichia coli, whether the expressed fusion protein can maintain the binding activity of PRRSV, and bind the smallest fragment of PRRSV What is it, what are the optimal binding and elution conditions, and can it be used for the concentration, purification and environmental monitoring of PRRSV?

Method used

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  • Self-polymerized peptide fused CD151 protein as well as preparation method and application thereof
  • Self-polymerized peptide fused CD151 protein as well as preparation method and application thereof
  • Self-polymerized peptide fused CD151 protein as well as preparation method and application thereof

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Embodiment Construction

[0034] Source of biological material:

[0035] 1. pET-30a vector: imported from Novagen, USA, and preserved in our laboratory.

[0036] 2. Escherichia coli DH5a: imported from BD Biosciences Clontech, USA, and preserved in our laboratory.

[0037] 3. BL21(DE3) Escherichia coli: imported from Novagen, USA, and preserved in our laboratory.

[0038] 4. PRRSV strain VR2332: caused by ATCC in the United States, preserved in our laboratory.

[0039] 5. Marc-145 cells: originated from ATCC in the United States and preserved in our laboratory.

[0040] 6. pET-IgV-CD151 vector: constructed in this laboratory (document: Zhang Yu, Sun Huaichang, Huang Yanyan, etc. Expression of pig CD151 recombinant antigen and its immune serum's PRRSV infection inhibitory effect. Journal of Yangzhou University (Agriculture and Life Science Edition), 2012, 33(2):1-5).

[0041] 7. ELK16-BoIFNg fusion protein: prepared in our laboratory (Document: Li Guangya. Expression, purification, activity detectio...

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Abstract

The invention belongs to the field of biotechnology research and in particular relates to self-polymerized peptide fused CD151 protein as well as a preparation method and application thereof. The self-polymerized peptide fused CD151 protein is composed of pig CD151 protein PRRSV (Porcine Reproductive and Respiratory Syndrome Virus) combined segment and an ELK16 self-polymerized peptide. The preparation method comprises the following steps: constructing an expression vector, expressing and purifying fused protein, determining a PRRSV combined segment and concentrating, purifying and detecting viruses. The self-polymerized peptide fused CD151 protein can be used for concentrating and purifying PRRSV and monitoring the environment. Compared with the other methods, the self-polymerized peptide fused CD151 protein has the advantages of being sensitive, simple, economical and the like when being used for the concentration, purification and PRRSV detection, can be used for concentrating and purifying the PRRSV of tissue samples and can also be used for PRRSV detection and separate culture of environment samples.

Description

technical field [0001] The invention relates to the field of biotechnology research, in particular to a preparation method of self-polypeptide fusion CD151 protein and its application in concentration, purification and environmental monitoring of porcine reproductive and respiratory syndrome virus. Background technique [0002] Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious and immunosuppressive infectious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), characterized by reproductive failure in sows, high mortality in young pigs, and respiratory Symptoms are characteristic and cause huge economic losses to the world pig industry. At present, PRRS is mainly controlled by vaccine immunization, but the immune protection effect of inactivated vaccine is not good, and live vaccine has potential safety hazards. Virus concentration and purification are important steps in the preparation of antibodies and vaccines. The exis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00C12N15/62C12N15/70C12N7/02
CPCC07K14/00C07K14/705C07K2319/00C12N7/00C12N2770/10051
Inventor 孙怀昌宗晓银李洋洋张鑫宇夏晓莉
Owner 江苏泰州动物药品工程技术中心
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