Fusion protein of beta 2-microglobulin and green fluorescent protein, preparation and use thereof
A technology of β2 microglobulin and green fluorescent protein, which is applied in the field of fusion protein of β2 microglobulin and green fluorescent protein, can solve the problems of time-consuming, laborious, low identification efficiency and high cost, and achieve accurate and reliable results and high identification efficiency , Easy and fast operation
Inactive Publication Date: 2009-07-15
ARMY MEDICAL UNIV
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In the past, the identification of CTL epitopes mainly used the method of synthesizing a large number of sequence misplaced repeat peptides and then screening them through lymphocyte experiments, which was time-consuming, laborious, expensive, and the identification efficiency was low.
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[0028] The amino acid sequence of the fusion protein β2M-ZsGreen of the present invention includes a first region having at least 85% identity to the sequence shown in SEQ ID No.1 and a first region having at least 85% identity to the sequence shown in SEQ ID No.2 The second region; wherein: the sequence shown in SEQ ID No.1 is the amino acid sequence of β2M composed of 102 amino acids, and the sequence shown in SEQ ID No.2 is the amino acid sequence of ZsGreen composed of 242 amino acids; in the art, Proteins with at least 85% sequence identity generally do not change the biological activity of the native protein; therefore, the fusion protein β2M-ZsGreen of the present invention includes a first region consisting of β2M or its derivative protein and a region composed of ZsGreen or its derivative protein. These derivative proteins include, but are not limited to: (1) proteins with one or more amino acid residues substituted, deleted or / and inserted, and the substituted or inse...
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The invention discloses fusion protein of beta2 microglobulin and green fluorescent protein. An amino acid sequence of the fusion protein comprises a first zone which has at least 85% homology with a sequence as shown in SEQ ID No.1 and a second zone which has at least 85% homology with a sequence as shown in SEQ ID No.2. The invention also discloses a gene encoding the fusion protein, a recombinant expression vector containing the gene, a method for preparing the fusion protein by the recombinant expression vector, and a method for detecting affinity between antigen peptide and MHC-I molecules by using the fusion protein. The detection method has the advantages of simple and rapid operation, accurate and reliable results, high identification efficiency, low cost, being applicable to CTL epitope identification with large flux and bright application prospect.
Description
technical field [0001] The invention relates to a fusion protein, in particular to a fusion protein of β2 microglobulin and green fluorescent protein, and also relates to a preparation method and application of the fusion protein. Background technique [0002] Cytotoxic T lymphocyte (CTL) epitopes are protein antigens processed by antigen-presenting cells (Antigen process cell, APC), and then combined with major histocompatibility complex-I (MHC-I) class molecules Combined with and finally presented to the T cell receptor (T cell receptor, TCR) for recognition, a short peptide that induces an effective immune response, generally 8 to 10 amino acids. CTL epitope identification has always been a hotspot in immunology, and is the key to epitope-based immunodiagnosis, immunotherapy and vaccine development. In the past, the identification of CTL epitopes mainly used the method of synthesizing a large number of sequence misplaced repeat peptides and then screening them through ly...
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IPC IPC(8): C07K19/00C12N15/62C12N15/63G01N33/53G01N15/00
Inventor 万瑛熊锐华王昊亮吴玉章
Owner ARMY MEDICAL UNIV