Production process of neurotrophic factor

A neurotrophic factor and production method technology, applied in the field of genetic engineering, can solve problems such as being unsuitable for large-scale preparation, and achieve the effects of shortening renaturation time and reducing polarization phenomenon

Inactive Publication Date: 2003-03-19
HANGZHOU JIUYUAN GENE ENG
View PDF0 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CNTF has been cloned and expressed by bacterial systems (Masiakowski, et al.1991, J.Neurosci.57: 1003-1012; Annalise Di Marco et al.1996, Proceedings of the National Academy of Scie

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
  • Production process of neurotrophic factor
  • Production process of neurotrophic factor
  • Production process of neurotrophic factor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0013] In order to improve the stability and biological activity of CNTF protein, compared with natural human CNTF, the primary structure has been changed, which is characterized in that the 17th cysteine ​​is changed to alanine (Cys 17 →Ala 17 ), the 63rd glutamic acid becomes arginine (Gln 63 →Arg 63 ), and remove the terminal 15 amino acids. At the same time, according to the characteristic that the codon preference of Escherichia coli is different from that of humans, the CNTF gene sequence was optimized and designed [see Figure 1]. Specifically, the codon corresponding to each amino acid in the CNTF natural gene sequence is transformed into a codon preferred by E. coli, while considering the G+C / A+T ratio balance of the entire gene sequence, and at the 5' end of the gene , 3' end and the middle are respectively introduced specific restriction sites, such as XhoI, EcoRI, AflIII to facilitate subsequent cloning and splicing. Based on the codon wobble theory, the CNTF ge...

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 present invention discloses the production process of neurotrophic factor. The production process includes computer-aided design, modifying and altering the gene of ciliary nerve trophic factor to obtain modified mutation human ciliary nerve trophic factor cDNA, artificially synthesizing the cDNA, constituting recombinant plasmid, converting to bacillus coli, high density expression in automatic fermentation tank; renaturation in tangent flow superfiltering mode; ion exchange chromatography, hydrophobic chromatography and molecular sieve chromatography to purify and thus to obtain high-purity ciliary nerve trophic factor. The present invention has the advantages of stable and efficient expression of mutant in bacillus coli, less polarization insuperfiltering course and high purity of renaturated ciliary nerve trophic factor.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a production method of neurotrophic factors. Background technique [0002] CNTF was first discovered in vitro to promote the survival of ciliary ganglion in chick embryos (Manthorpe et al., 1980, J.Neurochem.34:69-75). In the following decades, many more Related biological functions were discovered. Hughes et al. found that it can promote the differentiation of glial progenitor cells in the visual and cranial nerves of perinatal mice (Hughes et al., 1988, Nature 335:70-73), and also found that it can promote the differentiation of dorsal root ganglia in chick embryos. Growth (Skaper and Varon, 1986, Brain Res. 389:39-46). In addition, he can also promote the growth and differentiation of motor neurons, hippocampal neurons and parasympathetic spinal cord neurons (Ip, et al.1991, J.Neurosci.11: 3124-3134; Blottner, et al.1989, Neurosci.Lett .105:316-320)...

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): C07K14/435C07K14/48C12N15/12C12P21/02
Inventor 黄岩山王昌梅裘霁宛李辉孙瑛邹晔孙汉栋丁海
Owner HANGZHOU JIUYUAN GENE ENG
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