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DOI 10.13345/j.cjb.230114

Медицинская библиотека · Sheng wu gong cheng xue bao = Chinese journal of biotechnology · 2023

[Construction of foot-and-mouth disease virus like particles-induced expression vectors and screening of BHK-21 cell pools].

Tan Shuzhen, Dong Hu, Sun Shiqi, Guo Huichen

Аннотация на английскомСтатьяПеревод в очереди

Аннотация

Transient expression is the major method to express foot-and-mouth disease virus (FMDV) capsid proteins in mammalian cells. To achieve stable expression of FMDV capsid proteins and efficient assembly of virus like particles (VLPs) in cells, the plasmids of piggyBac (PB) transposon-constitutive expression and PB transposon-tetracycline (Tet) inducible expression vectors were constructed. The function of the plasmids was tested by fluorescent proteins. By adding antibiotics, the constitutive cell pools (C-WT, C-L127P) expressing P12A3C (WT/L127P) genes and the inducible cell pools (I-WT, I-L127P) expressing P12A3C (WT/L127P) genes were generated. The genes of green fluorescent protein, 3C protease and reverse tetracycline transactivator (rtTA) were integrated into chromosome, which was confirmed by fluorescence observation and PCR testing. The cell pool I-L127P has a stronger production capacity of capsid proteins and VLPs, which was confirmed by Western blotting and enzyme linked immunosorbent assay (ELISA), respectively. In conclusion, inducing the chromosomal expression of FMDV capsid proteins was firstly reported, which may facilitate the technical process of mammalian production of FMDV VLPs vaccine and the construction of mammalian inducible expression systems for other proteins.

Для цитирования: Tan Shuzhen, Dong Hu, Sun Shiqi, Guo Huichen. [Construction of foot-and-mouth disease virus like particles-induced expression vectors and screening of BHK-21 cell pools].. Sheng wu gong cheng xue bao = Chinese journal of biotechnology. 2023. DOI: 10.13345/j.cjb.230114.

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