SARS-CoV-2 coronavirus vaccine and preparation method thereof
A coronavirus, sars-cov-2 technology, applied in the field of biological genetic engineering, to achieve the effect of high biological activity, long-lasting immunogenicity and long half-life
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Embodiment 1
[0068] The construction of embodiment 1SARS-CoV-2 coronavirus vaccine Ad / S1
[0069] (1) Amplification of the S gene fragment:
[0070] Using the S gene obtained in the basic example as a template, amplify with the following primers:
[0071] V1 (SEQ ID NO: 1): TCC CCCGGG ATGTTCGTCTTCCTGGTCCT
[0072] V4 (SEQ ID NO: 4): CCC AAGCTT TTACCGGGCTCTTCTGGGAGAGT
[0073] The S1 gene is amplified, that is, the positions 1-2055 of the S gene of the basic example, and the amplified product is named S1.
[0074] (2) Construction of recombinant plasmids:
[0075] The S1 fragment of the PCR amplification product obtained above was identified by gel running and recovered by gel cutting, and digested with SmaI and HindIII at 37°C. Digest pShuttle with these two enzymes at the same time. Next, use T4 ligase to ligate the digested PCR product with pShuttle overnight at 16°C. The ligation product was transformed into Escherichia coli DH5α, positive clones were screened by ampicillin res...
Embodiment 2
[0078] The construction of embodiment 2SARS-CoV-2 coronavirus vaccine Ad / S1N
[0079] The difference with Example 1 is: the PCR amplification primers used in step (1) are:
[0080] V1 (SEQ ID NO: 1): TCC CCCGGG ATGTTCGTCTTCCTGGTCCT
[0081] V3 (SEQ ID NO: 3): CCC AAGCTT TTAGGCCACCTGGTTGCTTGTAT
[0082] The rest is the same.
[0083] The amplified product is named S1N fragment, and the final SARS-CoV-2 vaccine is named Ad / S1N vaccine, and the inserted gene is sequenced, and the sequencing results are shown in Figure 4 .
Embodiment 3
[0084] The construction of embodiment 3SARS-CoV-2 coronavirus vaccine Ad / S1C
[0085] The difference with Example 1 is: the PCR amplification primers used in step (1) are:
[0086] V2 (SEQ ID NO: 2): TCC CCCGGG ATGAGGGTGCAGCCAACCGAG
[0087] V4 (SEQ ID NO: 4): CCC AAGCTT TTACCGGGCTCTTCTGGGAGAGT
[0088] The rest is the same.
[0089] The amplified product is named S1C fragment, and the final SARS-CoV-2 vaccine is named Ad / S1C vaccine, and the inserted gene is sequenced, and the sequencing results are shown in Figure 5 .
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