SARS vaccine of adenovirus carrier and preparation method, application of coronavirus S gene
A coronavirus and vaccine preparation technology, applied in the field of biogenetic engineering, can solve problems such as infection and lack of vaccines
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Embodiment 1
[0035] The preparation method of adenovirus vector SARS vaccine:
[0036] The preparation of the adenovirus vector SARS vaccine is divided into two parts: the early construction and the later amplification:
[0037] Early build:
[0038] Get the SARS-related coronavirus spike(S F , S N , S M , S C ) gene, amplified by PCR method, after PCR, digested with Xbal+Kpnl 37°C, and simultaneously digested pShuttle with this enzyme, ligated, transformed into Escherichia coli DH5a, and used kanamycin (Kan R ) resistance screening positive clones, cultured and purified to obtain pS F / S N / S M / S C -Shuttle, use 1-Ceul+P1-Scel to digest, and use this enzyme to digest PAdeno-X TM , ligated after digestion, transformed into Escherichia coli DH5a, using ampicillin (Amp + ) resistance screening positive clones to obtain pAd-SF / S N / S M / S C .
[0039] Expand training:
[0040] get pAd-S F / S N / S M / S C Afterwards, after digestion with Pacl and linearization of the plasmi...
Embodiment 2
[0049] Cloned into the adenovirus vector is the N-terminal fragment of the S gene of SARS-associated coronavirus. All the other are with embodiment 1.
Embodiment 3
[0051] Cloned into the adenovirus vector is the middle fragment of the SARS-associated coronavirus S gene. All the other are with embodiment 1.
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