Recombinant canine sourced antibody scFv-Fc against H3N2 canine influenza virus
A technology of scfv-fc and VH-F is applied in the field of genetic engineering to achieve the effects of short preparation cycle, simple operation and mature production technology
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[0035] Example 1: Construction of pET-32a-scFv-Fc expression vector
[0036] 1 Virus strains, plasmids, cell lines and reagents
[0037] H3N2 subtype canine influenza virus strain A / Canine / Jiangsu / 06 / 2010 (H3N2) (06 for short), isolated and preserved by the laboratory, its GenBank sequence number is from JN247615 to JN247623. The strain and its sequence reference website: http: / / www.ncbi.nlm.nih.gov / nuccore. Rosetta (DE3) was purchased from Shanghai Weidi Biotechnology Co., Ltd. Escherichia coli DH5α competent cells and pClone007 Blunt Simple Vector Kit were purchased from Beijing Qingke Xinye Biotechnology Co., Ltd. (hereinafter referred to as Qingke). The pET-32a(+) prokaryotic expression vector is kept by the laboratory. Canine kidney cells (Madin-Darby canine kidney, MDCK) were purchased from the US ATCC and preserved by the laboratory.
[0038] Animal peripheral blood lymphocyte separation kit and TMB single-component chromogenic solution were purchased from Beijing S...
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