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Elastin-like polypeptide (ELP) and heat stress protein 90alpha (Hsp90alpha) fusion protein, and preparation method and application thereof

A heat stress protein, elastin-like technology, applied in biochemical equipment and methods, chemical instruments and methods, microorganism-based methods, etc., can solve problems to be studied, etc.

Active Publication Date: 2015-12-23
YANGZHOU UNIV
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Problems solved by technology

Our research group has established a simple and economical method for the concentration and purification of recombinant adenoviruses by using the fusion protein of ELP and adenovirus receptors. However, whether the fusion of ELP and Hsp90α or its fragments can maintain the reversible phase transition properties and be used for the concentration and purification of IBDV is still a question. to be studied

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  • Elastin-like polypeptide (ELP) and heat stress protein 90alpha (Hsp90alpha) fusion protein, and preparation method and application thereof
  • Elastin-like polypeptide (ELP) and heat stress protein 90alpha (Hsp90alpha) fusion protein, and preparation method and application thereof
  • Elastin-like polypeptide (ELP) and heat stress protein 90alpha (Hsp90alpha) fusion protein, and 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. pET / EI-GFP vector: imported from Princeton University, USA, preserved in our laboratory. Literature: FongBA, WoodDW. Expression and purification of ELP-intein-tagged target proteins in high cell density E. colifermentation. Microbial Cell Factories, 2010, 9:77.

[0037] 3. Escherichia coli DH5a: imported from BD Biosciences Clontech Company of the United States, and preserved in our laboratory.

[0038] 4. BLR (DE3) Escherichia coli: imported from Novagen, USA, and preserved in our laboratory.

[0039] 5. Chicken embryo fibroblast DF-1 line: imported from ATCC in the United States, preserved in our laboratory;

[0040] 6. Infectious bursal disease virus of Lukert strain: preserved by our laboratory (document: Lukert PD, Leonard J, Davis RB. Infectious bursal disease virus: antigen production and immunity. AmJ VetRes, 1975, 36 (4Pt2): 539...

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Abstract

The invention belongs to the field of biotechnology research, and particularly relates to an elastin-like polypeptide (ELP) and heat stress protein 90alpha (Hsp90alpha) fusion protein, and a preparation method and application thereof in infectious bursal disease virus (IBDV) concentration and purification. The ELP and Hsp90alpha fusion protein is composed of an IBDV binding segment M167a of ELP and Hsp90alpha. The preparation method comprises the following steps: construction of fusion expression vector, expression and purification of fusion protein, determination of IBDV binding segment, and concentration and elution of IBDV. Compared with other processes, the fusion protein for concentrating and purifying the IBDV is simple, and has the advantages of high speed and high economy. The prepared IBDV can be used for experimental research and vaccine preparation.

Description

technical field [0001] The invention relates to the field of biotechnology research, in particular to a preparation method of an elastin-like polypeptide and heat stress protein 90α fusion protein and its application in concentrating and purifying infectious bursal disease virus. Background technique [0002] Infectious bursal disease is a highly contagious infectious disease of chickens caused by infectious bursal disease virus (IBDV), which has caused huge economic losses to the world's poultry industry. Virus concentration and purification are important steps in virus molecular biology research and vaccine preparation. The existing IBDV concentration methods mainly include chloroform extraction and polyethylene glycol precipitation, etc., and the purification methods mainly include differential centrifugation, density gradient centrifugation and affinity chromatography. These methods are time-consuming, laborious, low recovery and high cost. shortcoming. In addition, th...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N7/02C12R1/93
Inventor 孙怀昌徐碧邵聪聪张鑫宇夏晓莉
Owner YANGZHOU UNIV
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