电穿孔
重组DNA
质粒
报告基因
生物
绿色荧光蛋白
基因
细胞生物学
植物
生物物理学
生物化学
基因表达
作者
Da‐Wei Li,Srinivasan Balamurugan,Jian-Wei Zheng,Wei‐Dong Yang,Jiesheng Liu,Hongye Li
出处
期刊:Methods in molecular biology
日期:2019-08-29
卷期号:: 175-179
被引量:4
标识
DOI:10.1007/978-1-4939-9740-4_19
摘要
Electroporation refers to the application of high strength electric pulse to create transient pores in the membrane, thereby enabling the passage of hydrophilic molecules into the cells. Based on the properties of cell and cell wall, the electroporation parameters vary among the algal species. Here, we demonstrated the optimized protocol for successful introduction of recombinant DNA (~5000 bp) into Nannochloropsis oceanica. The linearized recombinant plasmid that harbors eGFP and Bh-sle as the reporter and marker gene, respectively, was electroporated into the electrocompetent N. oceanica cells at voltage of 2200 V, 50 μF, resistance at 600 Ω using electroporator, and the transformed cells were then screened by molecular analysis. The report exemplifies a straightforward and reliable electroporation strategy for generating transgenic N. oceanica cells.
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