Rabies virus glycoprotein and nucleoprotein antigen epitope polypeptides, and screening and identification method and application thereof

A technology of rabies virus and antigenic epitope, applied in application, antiviral agent, chemical instrument and method, etc., can solve the problems of long time, difference, and sufficient rabies virus infection and incidence of rabies virus antibodies.

CN102924571AInactive Publication Date: 2013-02-13FUDAN UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2013-02-13
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention belongs to the technical field of biology, and particularly relates to screening and identification of antigen epitope polypeptides. The invention discloses screening, identification and application of a series of rabies virus glycoprotein and nucleoprotein antigen epitope polypeptides. The rabies virus glycoprotein and nucleoprotein are predicted by biological information means to obtain the candidate epitope polypeptides; and a lymphopoiesis experiment, ELISPOT experiment and a stream-type cell method are utilized to carry out in-vitro experimental verification on the subsequent epitope polypeptides to obtain the four rabies virus protein antigen epitope polypeptides. The invention is characterized in that the antigen epitope polypeptides respectively comprise a Th epitope and a CTL epitope, can stimulate the lymphopoiesis of the vaccine-immunized mouse in vitro and induce the cells to secrete related cell factors, and have the functions of killing virus-infected cells and stimulating the generation of the antibody. The invention can be used for developing rabies virus epitope vaccines and detecting the vaccine effect, and has important value for developing and producing immunologic function detection kits for rabies virus vaccines.
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Description

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a rabies virus glycoprotein and nucleoprotein epitope, a screening and identification method and application thereof. Background technique

[0002] Rabies is a relatively serious acute infectious disease that currently threatens human health. Once a human patient is bitten by a sick animal and becomes ill, the mortality rate is as high as 95%. The rabies virus belongs to the rhabdoviridae family (rhabdoviridae), lyssa virus (lyssa virus), and is bullet-shaped. Rabies virus can invade the human body through damaged skin or mucous membranes, and enter the central nervous system (CNS) of humans and all warm-blooded animals through nerve endings to cause rabies. The rabies virus genome nucleic acid is a single-stranded negative-strand RNA without segments, consisting of 11928 or 11932 nucleotides, with a relative molecular weight of about 4.6×10 6 , N, P, M, G, L genes are ...

Examples

Embodiment Construction

[0026] 1. Acquisition of rabies virus glycoprotein and protein amino acid sequence and epitope prediction

[0027] By searching the NCBI database, the full-length amino acid sequences of the rabies virus glycoprotein and nucleoprotein were obtained, numbered gi_32440976 and gi_294999043, respectively, and the specific sequences are shown in SEQ.ID.NO.5 and SEQ.ID.NO.6. CTL epitopes and Th epitopes were predicted for the sequences of rabies virus G protein and N protein obtained in the NCBI database. Mouse MHC class I molecules including H-2k d , H-2D d and H-2L d Three subclasses, MHC class II molecules include I-A d and I-E d Two subclasses. Therefore, in order to obtain CTL and Th epitopes, it is necessary to analyze and predict these subclasses respectively. For MHC class I molecules, SYFPEITHI and BIMAS software were used for prediction, and six sequences with higher common scores were selected, as shown in Table 1. For MHC-II molecules, RANKPEP software was used ...