Truncated type hepatitis c virus HCV NS3 antigen and preparation method and application thereof
A hepatitis C virus, truncated type technology, applied in the field of molecular biology, can solve the problems of inactivation of antigen and large protein, and achieve the effect of not easily missed detection
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Embodiment 1
[0033] Example 1: Obtaining of truncated HCVNS3 recombinant antigen protein sequence
[0034] According to the HCV sequences database (http: / / hepatitis.ibcp.fr.), the inventor screened out 6 NS3 antigen sequences belonging to 6 genotypes of HCV, and obtained the NS31252-1526aa truncated HCV NS3 amino acid sequence through sequence comparison and analysis. Search NCBI to obtain the corresponding nucleotide sequence, its amino acid sequence is shown in SEQ ID NO.1, and the corresponding nucleotide sequence is shown in SEQ ID NO.2. And further select the preferred codon of Escherichia coli, design and synthesize the nucleotide sequence of the truncated NS3 antigen gene, the codon nucleotide sequence is shown in SEQ ID NO.3, and the amino acid encoded by it corresponds to SEQ ID NO.2 The amino acid encoded by the nucleotide sequence.
Embodiment 2
[0035] Example 2: Cloning and expression of truncated HCVNS3 recombinant antigen
[0036] 1. Construction of truncated HCVNS3 recombinant antigen expression plasmid
[0037] 1.1 Synthesis of truncated HCVNS3 recombinant antigen
[0038] According to the truncated HCVNS3 antigen amino acid sequence (its amino acid sequence is shown in SEQIDNO.1), look up the NCBI related nucleotide sequence, obtain the corresponding nucleotide sequence (its nucleotide sequence is shown in SEQIDNO.2), use the large intestine The nucleotide sequence (such as SEQIDNO.3) obtained by modifying SEQIDNO.2 with Bacteria codon preference, entrusted Boshang Biotechnology Co., Ltd. to synthesize the nucleotides shown in the truncated HCVNS3 SEQIDNO.3, and respectively at the 5' end and Two restriction sites, BamHI and XhoI, were introduced into the 3' end to obtain the gene product.
[0039] 1.2 Construction of truncated HCVNS3 recombinant antigen expression plasmid
[0040] 1.2.1 Double digestion of t...
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