Expression of three reporter genes in four cell lines developed from Papilio demoleus Linnaeus (Lepidoptera: Papilionidae)

摘要

This paper used recombinant baculoviruses that carried three reporter genes, green fluorescent protein (GFP), beta-galactosidase, and secreted alkaline phosphatase (SEAP), to infect four new cell lines from Papilio demoleus Linnaeus larvae (named RIRI-PaDe-1, RIRI-PaDe-2, RIRI-PaDe-3, and RIRI-PaDe-4). The expression levels of the three recombinant proteins were detected at 24, 48, 72, 96, 120, and 144 h after infection and compared with Sf9 and High Five cells to evaluate the characteristics of these four cell lines as host cells. The inoculation densities of the tested cell lines were 2 x 10(4) cells/well (96-well plate) and 1 x 10(5) cells/well (24-well plate), and adding a volume of virus stock resulted in an MOI of 5.0. The results showed that the four cell lines could be infected by recombinant baculovirus and that cell lysis occurred 96 h after infection. In the four tested cell lines, only a small number of RIRI-PaDe-1 and RIRI-PaDe-3 cells expressed recombinant GFP and showed green fluorescence. The expression was much lower than that of Sf9 and High Five. Comparing the intracellular and extracellular activity of beta-galactosidase indicated that the P. demoleus cell system was more suitable for the expression of secreted proteins, and its extracellular beta-galactosidase level was close to that of Sf9, but the expression level of SEAP was far lower than those of Sf9 and High Five.