Recombinant Newcastle disease virus vector novel coronavirus vaccine candidate strain as well as construction method and application thereof
A technology for Newcastle disease virus and coronavirus, applied in the field of recombinant Newcastle disease virus vector new coronavirus vaccine candidate strain and its construction, can solve problems such as production risk, production capacity and cost barriers to large-scale popularization, and achieve low production cost and important Application value, the effect of highlighting the significance of public health security
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[0024] 1. Synthesis of the full-length S gene of the new coronavirus
[0025] According to the complete gene sequence (MN908947) of the Wuhan-Hu-1 strain published by Genebank, a C3756T silent mutation was introduced into the reading frame (ORF) of the S gene. The complete S gene ORF of the new coronavirus was synthesized by Nanjing Qingke Biotechnology Co., Ltd. The synthetic gene sequence is shown in SEQ ID NO.1.
[0026] 2. Insert the new crown S gene into the NDV genome
[0027] A viral reverse genetic vector based on artificial chromosome that can express NDV-LaSota full-length RNA ( figure 1 In A), the S gene of the new coronavirus was cloned into the Newcastle disease virus genome P-M by conventional homologous recombination, and a recombinant Newcastle disease virus vector containing the new coronavirus S gene was successfully constructed ( figure 1 in B). Homologous recombination using commercial kits: HD Cloning Kit (Clontech).
[0028] figure 1 Schematic dia...
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