生物
蛋白质表达
转座酶
细胞生物学
细胞培养
分子生物学
化学
基因
绿色荧光蛋白
转座因子
HEK 293细胞
质粒
重组DNA
转染
DNA
生物化学
遗传学
突变体
出处
期刊:Methods in Enzymology
日期:2021-01-01
卷期号:: 321-339
被引量:1
标识
DOI:10.1016/bs.mie.2021.06.016
摘要
Described here is the use of piggyBac transposase generated HEK293 stable cell pools for doxycycline-inducible protein production. The key benefits of the system are that low amounts of plasmid DNA are needed for transfection, high levels of protein expression can be achieved also for toxic proteins at robust scalability and reproducibility and the recombinant cell line can be stored as frozen cell bank. Transfection, selection, expression and purification of enhanced green fluorescence protein (eGFP) and SARS-CoV-2 Spike protein are described in this chapter.
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