Unlock instant, AI-driven research and patent intelligence for your innovation.

Construction of abrin A chain mutant and application as candidate vaccine antigen

A technology of vaccines and recombinant vectors, applied in antiviral agents, recombinant DNA technology, medical preparations containing active ingredients, etc.

Active Publication Date: 2013-07-10
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
View PDF1 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Chinese scholars have just started researching the detection and medical protection of abrin toxin biological warfare agents. There is no supporting diagnostic reagents and effective immunotherapy and protection methods for the terrorist attacks of abrin toxin. Therefore, it is urgent to establish and develop independent knowledge Property testing and protective equipment

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
  • Construction of abrin A chain mutant and application as candidate vaccine antigen
  • Construction of abrin A chain mutant and application as candidate vaccine antigen
  • Construction of abrin A chain mutant and application as candidate vaccine antigen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1, the acquisition of mutant mATA

[0032] 1. Obtaining of mutant mATA

[0033] According to the existing A-chain gene (rATA) sequence of abrin toxin (rATA nucleotide sequence is sequence 5, rATA amino acid sequence is sequence 6), using Quickchange Lighting Site-Directed Mutagenesis Kit (Stratagene Company, Catalog#210518) to carry out The mutant mATA-E164AR167L (plasmid) was obtained by site-directed mutagenesis. After sequencing, the gene contained in mATA-E164AR167L was named mATA. The 1-753rd nucleotide sequence of the terminal; the amino acid sequence of mATA encoded protein mATA is the sequence shown in Sequence 2 in the sequence list. Using Lasergene software to predict the antigenicity of the constructed mutants did not change significantly.

[0034] The mutation results were verified by sequencing, as follows:

[0035] The base A at the 491st position of sequence 5 is mutated to the base C at the 491st position of the sequence 1, and the base G a...

Embodiment 2

[0050] Embodiment 2, the biological activity detection of mutant mATA

[0051] 1. Quantification of Mutant Proteins

[0052] The purified mATA obtained in Example 1 was quantified using the BCA method protein quantification kit (purchased from Beijing Biotech Biotechnology Co., Ltd., Lot #20120613). The experiment was repeated three times, and the results were averaged.

[0053] Specific steps:

[0054] Kit composition: protein standard (5mg / mlBSA), Solution A, Solution B.

[0055] (1) When using, shake and mix Solution A. According to the number of samples, prepare an appropriate amount of BCA working solution with 50 volumes of Solution A plus 1 volume of Solution B (50:1), and mix well.

[0056] (2) Completely dissolve the protein standard (5mg / mlBSA), dilute it to 1.5mg / ml, 1.0mg / ml, 0.8mg / ml, 0.4mg / ml, 0.2mg / ml with PBS as the standard.

[0057] (3) Add the standard substance and the sample to be tested at each concentration mentioned above to the 96-well plate respect...

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

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
weightaaaaaaaaaa
Login to View More

Abstract

The invention discloses an abrin vaccine related protein and a coding gene and application thereof. The protein provided by the invention is named as mATA, and one of the following 1) and 2) is the protein: 1) protein consisting of amino acid residue sequences of a sequence 2 in a sequence table; and 2) protein consisting of amino acid residue sequences of the sequence 2 in the sequence table by substitution and / or deletion and / or addition of one or more amino acid residues, having the same functions and derived from the 1). Experiments prove that the mutant mATA obtained by point mutation isabrin A chain mutant protein obtained by recombinant expression; and by detection, the protein has the characteristics of low toxicity, no vascular leakage, good immunogenicity and the like, and can be used as a candidate vaccine antigen.

Description

technical field [0001] The invention relates to the construction of an abrin toxin A chain mutant and its use as a candidate vaccine antigen, in particular to a protein related to the abrin toxin vaccine, its coding gene and its application. Background technique [0002] With the rapid development of biotechnology and the development and utilization of biotoxins, the importance of toxins in weapons of mass destruction has been highlighted, and the threat of biotoxins and their bioterrorism is increasing day by day. Abrin toxin is composed of A and B chains, A chain is the effector chain, which has RNA N-glycosidase activity, and B chain is the binding chain, which mediates the A chain to enter the cell to play a toxic role. Acacia seeds from different origins contain abrin toxins with slight differences in the primary structure, which shows that their toxicity is different. The reported abrin toxin in the abdominal cavity of mice LD 50 Between 4-20μg / kg. Acacia seeds are w...

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 Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63A61K38/16A61P39/02
Inventor 王景林韩艳辉高姗康琳
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI