Laboratorio de Ingeniería Genética y Biología Celular y Molecular, Departamento de Ciencia y Tecnología, Universidad Nacional de Quilmes Roque Saenz Peña 352, CP B1876BXD Bernal, Argentina.
BMC Biotechnol. 2010 Sep 15;10:68. doi: 10.1186/1472-6750-10-68.
Anticarsia gemmatalis is a pest in South America's soybean crops, which could be controlled by the Multinucleopolyhedrovirus of A. gemmatalis (AgMNPV). Currently, its commercial production is based on infected larvae. However, the possibility of using modified baculoviruses in Integrated Pest Management programs has stimulated an interest to develop alternative multiplication processes. This study evaluated the AgMNPV production in UFL-Ag-286 cells previously deprived Fetal Bovine Serum.
Culture media containing 1% FBS during the previous 48 hours achieved a synchronized condition where 90% of cells were found in G0/G1 stage, showing the presence of non-filamentous actin. All characteristics were estimated from cellular viability tests, cell actin detection trials and flow cytometer cell cycle analysis. AgMNPV production was tested by transcript studies and budded viruses (BVs) and occlusion bodies (OBs) yield quantitation. Results showed that the productivity in FBS deprived cells was 9.8 times more in BVs and 3.8 times more in OBs with respect to non-treated cells.
UFL-Ag-286 cells previously deprived in FBS shown to be a better host for AgMNPV propagation, increasing the useful for both in vitro bioinsecticide production and applications such as recombinant protein expression or gene delivery.
美洲大豆的害虫 Agrius convolvuli 可被其多角体病毒(AgMNPV)控制。目前,其商业生产基于受感染的幼虫。然而,利用改良杆状病毒在综合虫害管理方案中的可能性激发了开发替代繁殖过程的兴趣。本研究评估了 UFL-Ag-286 细胞在先前缺乏胎牛血清(FBS)的情况下生产 AgMNPV。
在之前的 48 小时内含有 1% FBS 的培养基达到了同步条件,其中 90%的细胞处于 G0/G1 期,显示存在非丝状肌动蛋白。所有特征均来自细胞活力测试、细胞肌动蛋白检测试验和流式细胞仪细胞周期分析进行估计。通过转录研究和芽生病毒(BV)和包埋体(OB)产量定量测试 AgMNPV 的生产。结果表明,与未处理的细胞相比,在 FBS 剥夺细胞中的生产能力在 BV 中增加了 9.8 倍,在 OB 中增加了 3.8 倍。
先前在 FBS 中剥夺的 UFL-Ag-286 细胞显示出是 AgMNPV 繁殖的更好宿主,增加了体外生物杀虫剂生产和重组蛋白表达或基因传递等应用的用途。