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A fusion protein of elastin-like polypeptide and heat stress protein 90α, its preparation method and application

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: 2018-09-25
YANGZHOU UNIV
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  • Application Information

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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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  • A fusion protein of elastin-like polypeptide and heat stress protein 90α, its preparation method and application
  • A fusion protein of elastin-like polypeptide and heat stress protein 90α, its preparation method and application
  • A fusion protein of elastin-like polypeptide and heat stress protein 90α, its preparation method and application

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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: Fong BA, Wood DW. Expression and purification of ELP-intein-tagged target proteins in high cell density E. coli fermentation. Microbial Cell Factories, 2010, 9:77.

[0037] 3. Escherichia coli DH5a: imported from BD Biosciences Clontech, USA, 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. Lukert strain infectious bursal disease virus: preserved by our laboratory (document: Lukert PD, Leonard J, Davis RB. Infectious bursal disease virus: antigen production and immunity. Am JVet Res, 1975, 36 (4Pt 2): 539-540).

[0041] The ...

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Abstract

The invention belongs to the field of biotechnology research, and specifically relates to a fusion protein of an elastin-like polypeptide and a heat-stress protein 90α, a preparation method thereof, and an application in concentrating and purifying infectious bursal disease virus. The fusion protein of elastin-like polypeptide (ELP) and heat stress protein 90α (Hsp90α) is composed of ELP and the infectious bursal disease virus (IBDV) binding fragment M167a of Hsp90α. The preparation method includes construction of fusion expression vector, expression and purification of fusion protein, determination of IBDV binding fragment, concentration and elution of IBDV. Compared with other methods, using the fusion protein of the invention to concentrate and purify infectious bursal disease virus has the advantages of simplicity, speed and economy, and the prepared infectious bursal disease virus 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...

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Application Information

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Patent Type & Authority Patents(China)
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