Institute of Highland Forest Science, Chinese Academy of Forestry, Kunming, 650224, China.
Key Laboratory of Breeding and Utilization of Resource Insects, National Forestry and Grassland Administration, Kunming, 650224, China.
In Vitro Cell Dev Biol Anim. 2022 May;58(5):365-375. doi: 10.1007/s11626-022-00687-6. Epub 2022 Jun 2.
The baculovirus expression vector system using insect cells as a bioreactor has been used for in vitro expression of recombinant proteins and plays an important role in the fields of biology, agronomy, and medicine. Screening suitable host cell lines is an important part of the construction of insect cell baculovirus expression systems. In previous research, we used a single-cell cloning process with the Papilio xuthus cell line RIRI-PX1 and obtained the monoclonal cell line RIRI-PX1-C31. In this study, we compared the basic biological and recombinant protein expression characteristics of RIRI-PX1-C31 and its parent cell line RIRI-PX1 and found that the expression of recombinant β-galactosidase in RIRI-PX1-C31 was significantly higher than that in the parental cell line. Further serum-free adaptation studies confirmed that RIRI-PX1-C31 can adapt to the growth environment of Express Five Serum-free medium and that its expression level of recombinant β-galactosidase was significantly higher than that before adaptation.
利用昆虫细胞作为生物反应器的杆状病毒表达载体系统已被用于体外表达重组蛋白，在生物学、农学和医学领域发挥着重要作用。筛选合适的宿主细胞系是构建昆虫细胞杆状病毒表达系统的重要组成部分。在之前的研究中，我们使用 Papilio xuthus 细胞系 RIRI-PX1 的单细胞克隆过程，获得了单克隆细胞系 RIRI-PX1-C31。在本研究中，我们比较了 RIRI-PX1-C31 及其亲本细胞系 RIRI-PX1 的基本生物学和重组蛋白表达特性，发现 RIRI-PX1-C31 中重组β-半乳糖苷酶的表达明显高于亲本细胞系。进一步的无血清适应研究证实，RIRI-PX1-C31 可以适应 Express Five 无血清培养基的生长环境，其重组β-半乳糖苷酶的表达水平明显高于适应前。
