Process for producing recombinant insulin-like growth factor-1(IGF-1) amalgamation protein

A technology of recombinant human insulin and fusion protein, applied in the field of DNA recombination technology, can solve the problems of high cost, expensive enzyme preparation, unsuitable for large-scale production, etc., and achieve the effect of reducing purification cost, stable expression product and high-efficiency expression

Inactive Publication Date: 2009-05-13
FUJIAN MEDICAL UNIV
View PDF0 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the high price and high cost of enzyme preparations, this method is not suitable for large-scale production

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
  • Process for producing recombinant insulin-like growth factor-1(IGF-1) amalgamation protein
  • Process for producing recombinant insulin-like growth factor-1(IGF-1) amalgamation protein
  • Process for producing recombinant insulin-like growth factor-1(IGF-1) amalgamation protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0035] 1. Cloning of the fusion protein gene

[0036] 1.1 Design of tandem expression fusion protein

[0037] The first amino acid at the N-terminal of natural IGF-1 is Gly, not methionine. That is to say, the IGF-1 precursor synthesized in the human cytoplasm must be secreted outside the cell after the first methionine is removed. Therefore, if the IGF-1 gene containing the gene initiation code ATG is used to express alone, it is inconvenient to separate and purify on the one hand, and it is also very troublesome to remove methionine on the other hand. In the implementation, according to the characteristics that hydroxylamine can specifically crack the Asn-Gly peptide bond, and the first amino acid of natural IGF-1 is just Asn, and there is no Asn-Gly dipeptide sequence on the entire protein chain of IGF-1, in the design When designing an Asn-Gly sequence between the leader peptide and IGF-1, it promotes expression and facilitates purification.

[0038] 1.2 Gene primer des...

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 discloses a method for preparing recombined human insulin growth factor-1(IGF-1) fused protein, which belongs to the field of biological medicine field in biological technique. The invention adopts the genetic engineering fungus fermentation method for production: a. designing and synthesizing recombining human IGF-1 fused protein gene; b. constructing an expression vector of the recombined human IGF-1 fused protein; c. utilizing the expression vector to convert the host to construct the genetic engineering fungus; and d. utilizing the genetic engineering fungus to ferment the recombined human IGF-1 fused protein. In the method, according to the characteristics that the first amino acid of the natural IGF-1 is glycine(GLy, G), a leading peptide is added before the natural IGF-1, and a hydroxylamine specific cracking part(Asn-Gly peptide bond) is designed between the leading peptide and the natural IGF-1, thereby making the expression products stable and lowering purification cost. In the method, the fused protein in serial expression is used, wherein the N end is thioredoxin, and the C end is IGF-1, so that the expression products are more stable, and the separation method is simple and cheap. The method has wide application prospect.

Description

technical field [0001] The invention relates to the field of biomedicine in biotechnology, in particular to DNA recombination technology, that is, a method for constructing fusion protein of thioredoxin and human IGF-1 and preparing it by fermentation of genetically engineered coliform bacteria. Background technique [0002] In 1976, Rillderktercht isolated insulin-like growth factor-1 (insulin-like growth factor-1, IGF-1) from human serum, starting a new era of IGF-1 research. IGF-I has the functions of promoting growth and development and promoting substance substitution. It has attracted much attention in recent years. It has broad application prospects in the treatment of insulin-resistant diabetes, nerve damage, dwarfism, osteoporosis, and catabolic diseases. At present, IGF-1 has been industrialized in the treatment of diabetes and amyotrophic lateral sclerosis in foreign countries, but there is no product on the market in my country (there is a product with the same n...

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
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/62C12N15/12C12N15/70C12N1/21C07K19/00C07K14/65C12R1/19
Inventor 俞昌喜廖联明杨渐许盈
Owner FUJIAN MEDICAL 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