Expression of temperature responsive genes in cell cultures derived from Bombyx mori

Eun-Young Kim, M. Kang, K. Park, Kwangho Choi, S. Nho
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Abstract

Insects are heterotherms that exhibit a close relationship between their ecology (especially temperature changes) and physiology. In the present study, selected genes associated with cell death and temperature were examined to determine gene expression in Bombyx mori in high and low temperature environments. We determined the amount of dsRNA, different concentrations of dsRNA, and different type of cells to set the conditions most efficient for RNAi. We then prepared dsRNA transcripts of the genes associated with cell death and temperature response. We analyzed cell damage via Trypan blue staining and found that cell viability was reduced after knockdown of these genes. The special transduced cell lines produced in the present study can be applied in various research fields. We also expect that these cell lines can be used as a research tool for the precise functional analysis of various genes.
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家蚕细胞培养物中温度响应基因的表达
昆虫是异温动物,它们的生态(尤其是温度变化)和生理有着密切的关系。在本研究中,选择了与细胞死亡和温度相关的基因,检测了家蚕在高温和低温环境中的基因表达。我们确定了dsRNA的数量,不同浓度的dsRNA和不同类型的细胞,以设置最有效的RNAi条件。然后,我们制备了与细胞死亡和温度反应相关的基因的dsRNA转录本。我们通过台盼蓝染色分析细胞损伤,发现敲除这些基因后细胞活力降低。本研究产生的特殊转导细胞系可应用于各个研究领域。我们也期望这些细胞系可以作为研究工具,用于各种基因的精确功能分析。
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