Abstract
This paper used recombinant baculoviruses that carried three reporter genes, green fluorescent protein (GFP), β-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 × 104 cells/well (96-well plate) and 1 × 105 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 β-galactosidase indicated that the P. demoleus cell system was more suitable for the expression of secreted proteins, and its extracellular β-galactosidase level was close to that of Sf9, but the expression level of SEAP was far lower than those of Sf9 and High Five.
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Ding WF, Feng Y, Zhang X, Li X, Wang CY (2013) Establishment and characterization of a cell line developed from the neonate larvae of Papilio demoleus Linnaeus (Lepidoptera: Papilionidae). In Vitro Cell Dev. Biol.—Animal 49(2):108–113. https://doi.org/10.1007/s11626-012-9575-z
Ding WF, Feng Y, Zhang X, Li X, Wang CY, Xie SC (2015) Establishment and characterization of four cell lines developed from Papilio demoleus (Lepidoptera: Papilionidae). Acta Entomol Sin 58(8):826–835. https://doi.org/10.16380/j.kcxb.2015.08.002
Drugmand JC, Schneider YJ, Agathos SN (2012) Insect cells as factories for biomanufacturing. Biotechnol Adv 30(5):1140–1157. https://doi.org/10.1016/j.biotechadv.2011.09.014
Granados RR, Li GX, Derksen ACG, Mckenna KA (1994) A new insect cell line from Trichoplusia ni (BTI-Tn-5B1-4) susceptible to Trichoplusia ni single enveloped nuclear polyhedrosis virus. J Invertebr Pathol 64(3):260–266. https://doi.org/10.1016/S0022-2011(94)90400-6
Hink WF (1970) Established insect cell line from the cabbage looper, Trichoplusia ni. Nature 226(5244):466–467. https://doi.org/10.1038/226466b0
Murhammer DW (2016) Baculovirus and insect cell expression protocols (third edition). Humana Press, New York, p 26. https://doi.org/10.1007/978-1-4939-3043-2
Smith GE, Summers MD, Fraser MJ (1983) Production of human beta interferon in insect cells infected with a baculovirus expression vector. Mol Cell Biol 3(12):2156–2165. https://doi.org/10.1128/MCB.3.12.2156
Vaughn JL, Goodwin RH, Tompkins GJ, Mccawley P (1977) The establishment of two cell lines from the insect Spodoptera frugiperda (Lepidoptera; Noctuidae). In Vitro 13(4):213–220. https://doi.org/10.1007/BF02615077
Zhang X, Feng Y, Ding WF, Chen XM, Wang CY, Ma T (2011) Establishment and characterization of an embryonic cell line from Gampsocleis gratiosa (Orthoptera: Tettigoniidae). In Vitro Cell Dev Biol—Animal 47(4):327–332. https://doi.org/10.1007/s11626-011-9397-4
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This work was supported by the Fundamental Research Funds for the Central Non-profit Research Institution of CAF (No. CAFYBB2016ZD005). No third-party financial support for this work.
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Ding, WF., Zhang, X., Li, X. et al. Expression of three reporter genes in four cell lines developed from Papilio demoleus Linnaeus (Lepidoptera: Papilionidae). In Vitro Cell.Dev.Biol.-Animal 54, 194–199 (2018). https://doi.org/10.1007/s11626-017-0224-4
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DOI: https://doi.org/10.1007/s11626-017-0224-4