Meng Xiang-Qian, Zheng Gui-Ling, Zhao Chuan-De, Wan Fang-Hao, Li Chang-You
Key Lab of Integrated Crop Pest Management of Shandong Province, College of Agronomy and Plant Protection, Qingdao Agricultural University, Qingdao, Shandong, 266109, China.
State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China.
In Vitro Cell Dev Biol Anim. 2017 Aug;53(7):646-650. doi: 10.1007/s11626-017-0166-x. Epub 2017 Jun 29.
In this study, we describe a cell line, Ms-10C, cloned from the line QAU-Ms-E-10 (simplified Ms-10), an embryonic line from Mythimna separata. The cloned cell line was significantly more sensitive to nucleopolyhedrovirus (NPV). Ms-10C cells were mainly spherical with a diameter of 14.42 ± 2.23 μm. DNA amplification fingerprinting (DAF) confirmed the profile of PCR-amplified bands of the cloned cell line was consistent with those of the parental cell line, Ms-10. The sequencing result of the mitochondrial cytochrome c oxidase I (mtCO I) fragment confirmed that the amplified 636-bps mtCOI fragment was 100% identical to that of M. separata. Its chromosomes exhibited the typical characters of lepidopteran cell lines. Its population doubling time was 42.2 h at 27°C. Ms-10C was more sensitive than Ms-10 to both Autographa californica multiple nucleopolyhedrovirus (AcMNPV) and M. separata nucleopolyhedrovirus (MsNPV). At 4 d post infection, the infection rates of two viruses reached 94.2 and 92.3%, respectively. The availability of this cell clone strain will provide a useful tool for the basic research on nucleopolyhedrovirus and for potential application in expression of recombinant proteins with baculovirus expression vector system.
在本研究中,我们描述了一种从QAU-Ms-E-10细胞系(简称Ms-10,粘虫胚胎细胞系)克隆而来的细胞系Ms-10C。该克隆细胞系对核型多角体病毒(NPV)更为敏感。Ms-10C细胞主要呈球形,直径为14.42±2.23μm。DNA扩增指纹图谱(DAF)证实,克隆细胞系PCR扩增条带的图谱与亲本细胞系Ms-10一致。线粒体细胞色素c氧化酶I(mtCO I)片段的测序结果证实,扩增得到的636-bps mtCOI片段与粘虫的该片段100%相同。其染色体表现出鳞翅目细胞系的典型特征。在27°C条件下,其群体倍增时间为42.2小时。Ms-10C对苜蓿银纹夜蛾多粒包埋核型多角体病毒(AcMNPV)和粘虫核型多角体病毒(MsNPV)均比Ms-10更为敏感。感染后4天,两种病毒的感染率分别达到94.2%和92.3%。该细胞克隆株的获得将为核型多角体病毒的基础研究以及杆状病毒表达载体系统在重组蛋白表达中的潜在应用提供有用工具。