Rapid and high efficiency transformation of Chlamydomonas reinhardtii by square-wave electroporation
Rapid and high efficiency transformation of Chlamydomonas reinhardtii by square-wave electroporation
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方波电穿孔快速高效转化莱茵衣藻
DOI：
10.1042/bsr20181210
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发表时间：
2018-12
影响因子：
4
通讯作者：
Wen Wang
中科院分区：
文献类型：
--
作者：
Liang Wang;Lijing Yang;Xin Wen;Zhuoya Chen;Qiaoying Liang;Jialing Li;Wen Wang
莱茵衣藻是一种单细胞绿藻，是研究各种生理过程和生物工业应用的模式生物。为了探索生理过程的分子机制或建立工程细胞系，需要将外源DNA整合到基因组中进行插入突变或转基因表达。然而，现有的构建突变体文库的电穿孔方法并没有认真考虑选择标记DNA的数量。在这里，我们报道了一种利用方波电穿孔系统对细胞壁菌株进行快速高效转化的技术。这种电穿孔方法的最终产量为每μg外源DNA2-6×10~3个转化子，且以菌株依赖的方式进行。总的来说，这种电穿孔技术是建立突变体库以筛选感兴趣的表型的简单和适用的方法。
Chlamydomonas reinhardtii, the unicellular green algae, is the model organism for studies in various physiological processes and for bioindustrial applications. To explore the molecular mechanisms underlying physiological processes or to establish engineered cell lines, the exogenous DNA needs to be integrated into the genome for the insertional mutagenesis or transgene expression. However, the amount of selected marker DNA is not seriously considered in the existing electroporation methods for mutants library construction. Here, we reported a rapid-and-high-efficiency transformation technique for cell-walled strains using square-wave electroporation system. The final yield with this electroporation method was 2–6 × 103 transformants per μg exogenous DNA for cell-walled strains in a strain-dependent manner. In general, this electroporation technique was the easy and applicable way to build a mutant library for screening phenotypes of interest.
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影响因子：
2.6
作者：
Pratheesh, P. T.;Vineetha, M.;Kurup, G. Muraleedhara
通讯作者：
Kurup, G. Muraleedhara
构建衣藻正向和反向遗传学的多用途插入突变体库
影响因子：
5.1
作者：
Cheng X;Liu G;Ke W;Zhao L;Lv B;Ma X;Xu N;Xia X;Deng X;Zheng C;Huang K
通讯作者：
Huang K
影响因子：
--
作者：
J. A. Nelson;P. Lefebvre
通讯作者：
J. A. Nelson;P. Lefebvre
影响因子：
4.6
作者：
Kang S;Kim KH;Kim YC
通讯作者：
Kim YC
影响因子：
3.3
作者：
Kosuke Shimogawara;S. Fujiwara;A. Grossman;H. Usuda
通讯作者：
Kosuke Shimogawara;S. Fujiwara;A. Grossman;H. Usuda