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Establishing and expressing method for chimeric vector of EV71 neutralization epitope and norovirus P structure domain

An antigenic epitope, EV71 technology, applied in the field of genetic engineering, can solve the problems of complex production process and high production cost, and achieve the effects of simple preparation and simple purification process

Pending Publication Date: 2016-12-14
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

For enterovirus EV71, although its inactivated vaccine has been approved for marketing, its production process is complicated, the production cost is high, and there are certain risks

Method used

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  • Establishing and expressing method for chimeric vector of EV71 neutralization epitope and norovirus P structure domain
  • Establishing and expressing method for chimeric vector of EV71 neutralization epitope and norovirus P structure domain
  • Establishing and expressing method for chimeric vector of EV71 neutralization epitope and norovirus P structure domain

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Experimental program
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Embodiment Construction

[0045] (1) Construction and identification of recombinant plasmids

[0046] Plasmid P42 is from Cincinnati Children's Medical Research Center, USA (patent protected, patent number: WO2010144602-A2; US2010322962-A1; WO2010144602-A3; EP2440582-A2; CN102612525-A; JP2012529298-W; HK1169426-A4; US841 ; US2013273090), it is a vector containing the GST tag and human norovirus GII.4 genotype P domain (GI: 146387016) gene, the plasmid map see figure 1 . The amino acid sequences of the neutralizing antigenic epitopes of EV71 were obtained from literature reports, namely the SP55 (163aa-177aa) and SP70 (208aa-222aa) on VP1 and the SP28 (136-150aa) epitopes on VP2 ( figure 2 ), optimize the nucleotide sequence of each single epitope according to the preference of Escherichia coli codon expression, and design the tandem epitope composed of single epitope combination: SP55-SP70-SP28, SP55-SP70, SP55-SP28, SP70-SP28 (the nucleotide sequences are respectively shown in SEQ ID NO: 2 to SEQ I...

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Abstract

The invention aims at providing chimeric protein of enterovirus EV71 neutralization epitope and a human norovirus P structure domain and an establishing method of a chimeric vector. By means of immunoblotting, it is verified that the chimeric protein of the enterovirus EV71 neutralization epitope and the norovirus P structure domain can be used for research and development of detection kits of EV71 and norovirus and novel bivalent subunit vaccines.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and relates to the construction and expression of EV71 neutralizing antigenic epitope and norovirus P domain chimeric protein and its expression vector construction method, which can be used for the detection kits of enterovirus EV71 and norovirus and Development of novel bivalent subunit vaccines. Background technique [0002] Acute gastroenteritis is a serious public health problem at present. It not only affects people's life, work and combat effectiveness of troops, but also can cause death in severe cases, especially in children and immunocompromised populations. Norovirus diarrhea is prevalent worldwide. Enterovirus 71 (EV71) is the most common pathogen in hand, foot and mouth patients. The virus mainly infects children, causing a variety of neurological symptoms and seriously affecting children's health, growth and development. Since the 1990s, the prevalence of EV71 has show...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N15/70C12N15/66C07K1/36C07K1/34C07K1/22
Inventor 刘红旗李超刘波陶玉芬李昕潼刘建生
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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