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Blue-green algal virus protein N mutant, modified derivative and application thereof

A technology of cyanobacteria virus and mutants, applied in the field of cyanobacteria virus protein N mutants, can solve the problems of loss of anti-HIV activity and low activity, and achieve the effect of enhanced antiviral activity

Inactive Publication Date: 2010-02-03
JINAN UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the anti-HIV activity of CVN (Q62C) is lower than that of unmutated CVN, and since the selected modification site is inside CVN, the body modified by mPEG-maleimide 30KDa (mPEG-MAL-30KDa) Variant nearly loses activity against HIV

Method used

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  • Blue-green algal virus protein N mutant, modified derivative and application thereof
  • Blue-green algal virus protein N mutant, modified derivative and application thereof
  • Blue-green algal virus protein N mutant, modified derivative and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0053] 1. Construction of recombinant plasmid pET3c-6His-SUMO-LCVN

[0054] The construction and synthesis of the SUMO-L-CVN gene is carried out in two steps. First, the L-CVN gene is synthesized by two PCRs. The first PCR uses the pET3c-SUMO-CVN plasmid as a template, and F1-CVN and R-CVN as upstream and downstream primers. The reaction system was 1 ng template, 1 μM upstream and downstream primers, 20 μl Taq PCR MasterMix, added water to 40 μl, the reaction mixture was denatured at 94 °C for 1 min, annealed to 55 °C, maintained for 1 min, extended at 72 °C for 1 min, and performed 29 cycles. The reaction product was subjected to 1% agarose gel electrophoresis, and the target fragment was recovered from the gel, which was used as a template for the next round of PCR. In the second PCR, the PCR product of the previous round was used as a template, and F2-CVN and R-CVN were used as upstream and downstream primer pairs (the F2-CVN primer contained a flexible polypeptide encodin...

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Abstract

The invention belongs to the field of biological medicines, in particular to a blue-green algal virus protein N mutant, a PEG modified derivative and application thereof. The blue-green algal virus protein N mutant comprises two sequences, one is connecting peptide with hydrophilic flexibility, and the other is a blue-green algal virus protein N sequence optimized by trinucleotide. The derivativeis obtained by PEG modification of the blue-green algal virus protein N mutant; the mutant and the derivative thereof show favorable anti-HIV activity. The invention is applied to preparing anti-HIV or other virus microorganisms.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a cyanobacteria viral protein N (CVN) mutant, its PEG-modified derivative and their application in pharmacy. Background technique [0002] Cyanovirin-N (CVN) is a protein with anti-HIV activity that can be extracted from cyanobacteria by American scientists BOYD et al. CVN can specifically bind to the capsid protein gp120 of human immunodeficiency virus HIV-1 with high affinity to exert antiviral activity. Broad spectrum and stable properties make CVN protein a valuable antiviral drug [BOYD M R, GUSTAFSON K R, MCMAHON J B, et al. protein that binds viral surface envelope glycoprotein gp 120: Potential applications to microbicide development [J]. Antimicrobial Agents and Chemotherapy, 1997, 41(7): 1521-30.]. However, because the molecular weight of the protein is small and there are two disulfide bonds in the molecule, it is difficult to express the protein in Escherichia ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/405C12N15/31C12N15/63C12N1/21C07K17/08A61K38/16A61K47/48A61P31/18C12R1/89C12R1/19
CPCA61K47/48215A61K38/00C07K14/195A61K47/60A61P31/18
Inventor 熊盛陈伟钱垂文王一飞北里海雄
Owner JINAN UNIVERSITY
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