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. 2023 May 4;11(5):941. doi: 10.3390/vaccines11050941

Figure 1.

Figure 1

The construction and formation of LV20. (A) The construct of pFastBacDual-MH-L20. The functional domains found in HBHA can be divided into transmembrane domain (1–18), coiled-coil domain (19–99), and C-terminal domain (100–199). The functional domains found in MTP can be divided into inner membrane region (1–8), transmembrane region (9–26), and outer membrane region (27–103). The linked outer membrane region of HBHA and MTP (L20 in blue) were inserted between the transmembrane region and extracellular signal peptide of HA (represented in yellow) to construct the recombinant double expression plasmid pFastBacDual-MH-L20. M1, influenza matrix protein; pP10, P10 promoter; SP, extracellular signal peptide of HA; and TM-CT, transmembrane-cytoplasmic tail of HA. (B) The recombinant gene expression and formation of virus-like particles. The recombinant double expression plasmid pFastBacDual-HM-L20 was transformed into DH10 Bac™ E. coli (Invitrogen, Carlsbad, CA, USA) strain, and positive clones contained the recombinant bacmid -HM-L20 were obtained by antibiotic and blue–white screening. The correct recombinant bacmid DNA was extracted and transfected into Sf9 cells to obtain recombinant baculovirus. Virus-like particles were produced by recombinant baculovirus expression system.