In this paper, four established cell lines derived from newly hatched larvae of Papilio demoleus Linnaeus and 57 single-cell clones derived from the 3 lines were used as test materials. Recombinant β-galactosidase baculovirus AcMNPV-Gal was used to infect the P. demoleus L. cell lines and the single-cell clones, and recombinant protein expression in each cell line was detected and compared. Three clonal cell lines, RIRI-PaDe-1-C1, RIRI-PaDe-2-C6 and RIRI-PaDe-3-C52, which showed significantly higher β-galactosidase expression levels than those of the parental cell lines, were selected. Five types of commercial serum-free media for insect cells, Express Five SFM, Ex-Cell 405, Sf-900III SFM, Sf-900II SFM and HyClone Serum-Free Media, were used to adapt RIRI-PaDe-2-C6 cells and RIRI-PaDe-3-C52 cells to serum-free culture conditions, and the growth characteristics of the cells and the exogenous protein expression characteristics before and after adaptation were compared. The results showed that RIRI-PaDe-2-C6 cells could stably proliferate in Ex-Cell 405, RIRI-PaDe-3-C52 cells could stably proliferate in Express Five SFM and Ex-Cell 405, and the rate of proliferation of and the level of expression of β-galactosidase in RIRI-PaDe-3-C52 cells were significantly increased in Express Five SFM.

在本文中，使用来自新孵化的凤蝶幼虫的四个已建立的细胞系和来自这三个系的57个单细胞克隆作为测试材料。使用重组β-半乳糖苷酶杆状病毒AcMNPV-Gal感染蠕形疟原虫细胞系和单细胞克隆，检测并比较每种细胞系中的重组蛋白表达。三种克隆细胞系RIRI-PaDe-1-C1，RIRI-PaDe-2-C6和RIRI-PaDe-3-C52显示出明显更高的β选择比亲代细胞系的β-半乳糖苷酶表达水平。用于昆虫细胞的五种商业无血清培养基，快速五型SFM，Ex-Cell 405，Sf-900III SFM，Sf-900II SFM和HyClone无血清培养基用于适应RIRI-PaDe-2-C6细胞和比较了RIRI-PaDe-3-C52细胞在无血清条件下的生长情况，以及适应前后细胞的生长特性和外源蛋白表达特性。结果表明，RIRI-PaDe-2-C6细胞可以在Ex-Cell 405中稳定增殖，RIRI-PaDe-3-C52细胞可以在Express 5 SFM和Ex-Cell 405中稳定增殖，其增殖速率和增殖速率均高于正常细胞。在Express 5 SFM中，RIRI-PaDe-3-C52细胞中β-半乳糖苷酶的表达水平显着增加。