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美洲大蠊(Periplaneta americana L.)胚胎细胞系对 AcMNPV 表现出易感性。 (蜚蠊目:蜚蠊科)

An embryonic cell line from the American cockroach Periplaneta americana L. (Blattaria: Blattidae) exhibits susceptibility to AcMNPV.

机构信息

Key Laboratory of Breeding and Utilization of Resource Insects of National Forestry and Grassland Administration, Institute of Highland Forest Science, Chinese Academy of Forestry, Kunming, Yunnan Province, 650224, China.

出版信息

In Vitro Cell Dev Biol Anim. 2022 Apr;58(4):278-288. doi: 10.1007/s11626-021-00628-9. Epub 2022 Apr 22.

Abstract

Although the baculovirus expression vector system (BEVS) is widely used in the production of recombinant proteins, only a few lepidopteran insect cell lines have been successfully used so far. This study aimed at evaluating the characteristics of an embryonic cell line from the American cockroach Periplaneta americana L., RIRI-PA1, and determining whether it could be used in recombinant protein expression. Wild type Autographa californica multiple nucleopolyhedrovirus (AcMNPV-wt) and green fluorescent protein (GFP)-replicating recombinant baculoviruses (AcMNPV-GFP) were used to infect RIRI-PA1 respectively, demonstrating that RIRI-PA1 cells could be infected by AcMNPV and express recombinant proteins. Within 24 h of infection with AcMNPV-GFP, the GFP expression was higher than that in Sf21 cells. Furthermore, the infection of RIRI-PA1 cells increased gradually (multiplicity of infection, 10) within 24 h, while in Sf21 cells, the infection only began to increase within 48 h. However, after exposure for 96-168 h, the virus progeny and recombinant protein production of RIRI-PA1 cells was lower than those of Sf21 cells. Western blotting revealed that RIRI-PA1 cells could express recombinant GFP, and the protein expression level positively correlated with the multiplicity of infection. In conclusion, this is the first report that a cell line from P. americana has shown susceptibility to infection from a baculovirus and likewise express recombinant protein. Although the yield of recombinant GFP was not as high as that of Sf21 cells, the results nonetheless showed that RIRI-PA1 had an infection rate advantage in the short term (within 24 h of infection), which is of great value for further development and utilization.

摘要

尽管杆状病毒表达载体系统(BEVS)被广泛应用于重组蛋白的生产,但迄今为止,仅有少数鳞翅目昆虫细胞系成功应用于该系统。本研究旨在评估美洲大蠊(Periplaneta americana L.)胚胎细胞系 RIRI-PA1 的特性,并确定其是否可用于重组蛋白表达。野生型苜蓿银纹夜蛾核型多角体病毒(AcMNPV-wt)和绿色荧光蛋白(GFP)复制型重组杆状病毒(AcMNPV-GFP)分别感染 RIRI-PA1,结果表明 RIRI-PA1 细胞可被 AcMNPV 感染并表达重组蛋白。感染 AcMNPV-GFP 后 24 h,GFP 表达水平高于 Sf21 细胞。此外,感染 RIRI-PA1 细胞的感染率在 24 h 内逐渐升高(感染复数,10),而 Sf21 细胞的感染仅在 48 h 内开始增加。然而,暴露 96-168 h 后,RIRI-PA1 细胞的病毒子代和重组蛋白产量均低于 Sf21 细胞。Western blot 分析显示,RIRI-PA1 细胞可表达重组 GFP,且蛋白表达水平与感染复数呈正相关。综上所述,本研究首次报道美洲大蠊细胞系对杆状病毒具有易感性,并能表达重组蛋白。尽管重组 GFP 的产量不如 Sf21 细胞高,但结果表明 RIRI-PA1 在短期内(感染后 24 h 内)具有较高的感染率,这对于进一步的开发和利用具有重要价值。

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