Tobacco etch virus protease active inclusion body as well as preparation method and application thereof

A technology of tobacco etch virus and protease activity, applied in the biological field

Inactive Publication Date: 2016-08-24
YANGZHOU UNIV
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this strategy is easy to think of, it faces the following technical problems in the specific design: how to construct an N-terminal self-polypeptide fusion expression vector, whether the natural TEVP coding seque

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
  • Tobacco etch virus protease active inclusion body as well as preparation method and application thereof
  • Tobacco etch virus protease active inclusion body as well as preparation method and application thereof
  • Tobacco etch virus protease active inclusion body as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0033] Source of biological material:

[0034] 1. pET-30a vector: imported from Novagen, USA, and preserved in our laboratory.

[0035] 2. Escherichia coli DH5a: imported from BD Biosciences Clontech, USA, and preserved in our laboratory.

[0036] 3. BL21(DE3) Escherichia coli: imported from Novagen, USA, and preserved in our laboratory.

[0037] 4. MDBK cells: imported from ATCC in the United States and preserved in our laboratory.

[0038] 5. Vesicular stomatitis virus (VSV): preserved by our laboratory (document: Xu Yaoxian, Zhou Xiaofeng, Liu Lide. Molecular Virology. Wuhan: Hubei Science and Technology Press, 2000.300-302).

[0039] 6. pGEX-BoIFNg vector: Constructed and preserved by our laboratory (document: Xu Jinjun, Qin Aijian, Jin Wenjie, etc. Cloning of cow gamma interferon gene and its expression in Escherichia coli. Journal of Yangzhou University, 2003, 24(1): 5-10).

[0040] The specific operation steps are as follows:

[0041] 1. Construction of pP16P-TEVP ...

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 belongs to the field of biotechnology research, and particularly relates to a TEVP (Tobacco Etch Virus Protease) active inclusion body as well as a preparation method thereof and application thereof in the preparation of a recombinant protein. The TEVP active inclusion body consists of an ELK16 self-polymeric peptide and TEVP that are separated by two PT connectors. The preparation method of the TEVP active inclusion body comprises the steps of an expression vector construction, an active inclusion body expression and purification, recombinant protein cleavage and target protein recovery. Compared with existing TEVP, the TEVP active inclusion body proviced by the invention has the advantages of simple preparation, easiness for being removed from an end product, low cost and the like, and can be applied to prepare various recombinant proteins.

Description

technical field [0001] The invention relates to the field of biotechnology research, in particular to a preparation method of tobacco etch virus protease activity inclusion body and its application in the preparation of recombinant protein. technical background [0002] Purification of recombinant proteins has always been one of the bottlenecks restricting molecular biology research and production of biological products. Gene fusion technology can simplify recombinant protein purification, and the most commonly used fusion tag is an affinity tag. Although affinity tag fusion proteins can be purified by chromatography, there are disadvantages such as time-consuming, expensive and difficult to scale up. In most cases, these fusion proteins require removal of the affinity tag in order for the protein of interest to fold properly or become active. A common practice is to introduce a protease recognition site between the fusion tag and the target protein, and then use protease ...

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
IPC IPC(8): C12N9/50C12N15/70C12N15/57C12P21/06
CPCC12N9/506C07K14/57C12N15/70C12N2800/101C12N2800/22C12P21/06
Inventor 孙怀昌李光亚卢会鹏肖真真李洋洋周晓慧谭笑张鑫宇夏晓莉
Owner YANGZHOU UNIV
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