Series connection expression method from virus macrophage inflammatory protein IIN end 15 peptide

A technology of macrophages and inflammatory proteins, applied in antiviral agents, peptide/protein components, botanical equipment and methods, etc., can solve problems that have not occurred before

Inactive Publication Date: 2007-03-14
广州法蕊仕药业科技有限公司
View PDF0 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there have been no related reports on the treatment of AIDS from viral macrophage inflammatory protein II N-terminal 15 peptides.

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Firstly, the gene sequence of the N-terminal 15 peptide was found from the mature viral macrophage inflammatory protein II gene,

[0040] 5'ctg gga gcg tcc tgg cat aga ccg gac aag tgc tgt ctc ggt tac 3'

[0041] The amino acid sequence is: LGASWHRPDKCCLGY.

[0042] According to the yeast's codon preference, the optimized sequence is ttg ggt gct tcctgg cac aga cca gac aag tgc tgt ttg ggt tac, and the nucleic acid sequence of the reverse amino acid sequence is: tac ggt ttg tgt tgc aag gac cca aga cac tgg tcc gct ggt ttg, reverse amino acid sequence: YGLCCKDPRHWSAGL.

[0043] Thrombin is selected as the protease, and the amino acid sequence of its digestion is L-V-P-R↓G-S The restriction site has been marked, and the nucleic acid sequence is: CTG GTC CCC CGG GGC AGC optimizes it according to the yeast's preference for codons, and the optimized sequence For, TTG GTC CCA AGA GGT TCC.

[0044] The gene sequence designed and synthesized is:

[0045] CCGGAATTC TTG GTC CCA A...

Embodiment 2

[0061] Design the synthetic gene as follows: the plasmid used is pTYB1

[0062] CTAGCTAGC ttg ggt gct tcc tgg cac aga cca gac aag tgc tgt ttg ggttac gct gct gct gct ttg ggt gct tcc tgg cac aga cca gac aag tgc tgt ttgggt tac gct gct gct gct tac ggt ttg tgt tgc aag gac cca aga cac tgg tccgct ggt ttg CTCGAGGAC....tag (self-breaking)

[0063] Synthesize the target gene according to the above gene sequence, and construct the recombinant plasmid through gene recombination

[0064] pTYB1-target gene, the recombinant plasmid was transformed into Escherichia coli DH5a, screened with LB plates containing penicillin (100ug / ml), and the single colony of Escherichia coli obtained from the screening was amplified and cultured, and the plasmid was extracted from it for enzyme digestion and sequencing identification, and the screening was positive clone.

[0065] Cultivate positive bacteria in LB medium (containing penicillin 100ug / ml) at a temperature of 37°C, and induce expression with IP...

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 genetic engineering tandem expression process of pentadecapeptide originated from virus macropage inflammatory protein II N terminal includes: synthesizing one gene segment including several pentadecapeptides connected through protein cleavage sequence gene by using virus macropage inflammatory protein II N terminal pentadecapeptided as target and in a eukaryotic expression system; adding limiting cleavage sites to the 5' and 3' terminals of the gene segment; connecting the gene segment to plasmid vector by means of gene recombination, converting saccaromycete to screen, express and purify target protein; cleaving the purified protein with protease, and separating and purifying to obtain target polypeptide. The polypeptide can antagonize CXCR4 pentadecapeptide, and has the function of preventing and treating AIDS and wide medicine application foreground.

Description

technical field [0001] The invention relates to the pharmaceutical field of medical gene recombination synthesis, in particular to a tandem expression method of 15 peptides derived from the N-terminus of viral macrophage inflammatory protein II. Background technique [0002] AIDS (AIDS) is a kind of human immunodeficiency virus (human immunodeficiency virus, referred to as HIV) invades the human body and destroys the immune function of the human body, causing various incurable infections and tumors in the human body, and finally leading to the death of the infected person. a serious infectious disease. It spreads rapidly among the population mainly through blood, sexual contact, mother-to-child transmission, etc., but lacks effective preventive vaccines and therapeutic drugs, and is considered to be the plague of the 21st century. The latest research finds that Chinese people are more susceptible to HIV infection than whites and blacks, making us face an even bigger challen...

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): C12N15/33C12N15/62A61K38/17A61P31/18
Inventor 叶石敦赵宝梅
Owner 广州法蕊仕药业科技有限公司
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