红豆杉
继代培养(生物学)
DNA甲基化
紫杉醇
羟基烷酸
细胞周期
扩增片段长度多态性
甲基化
生物
DNA
化学
细胞
植物
生物化学
遗传学
医学
细菌
癌症
基因
人口
基因表达
环境卫生
遗传多样性
作者
Liqin Li,Xiaoli Li,Chunhua Fu,Chunyu Zhao,Liang Yu
标识
DOI:10.1016/j.procbio.2013.01.013
摘要
Yields of paclitaxel decreased with repeated subculturing of Taxus media cells. We used minimal growth conservation and manipulation of genome methylation to sustain paclitaxel production by Taxus media cell cultures. The subculture period of Taxus cells can be prolonged to 180 d by incubating them at a low temperature (5 °C). Paclitaxel levels increased in the cells after conservation and during the first recovery subculture cycle, and then decreased during the subsequent recovery subculture cycle. Analysis of genetic variations in these cultures using amplified fragment-length polymorphism (AFLP) technology identified only two polymorphic bands associated with the second and sixth recovery cycle cultures. However, the results of high-performance liquid chromatography indicated that DNA methylation increased during the course of repeated subculturing. A decrease in DNA methylation level caused by treatment with 5-Aza-2′-deoxycytidine coincided with an increase in paclitaxel levels. Simultaneous exposure to both methyl jasmonate and 5-Aza-2′-deoxycytidine increased paclitaxel levels to 320.43 μg g−1 (dry weight), which is more than six times the paclitaxel content before conservation. To our knowledge, this is the first report about improving paclitaxel production by ensuring sustainable use of Taxus cells.
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