Paris-Saclay University, Orsay, France.
Methods Mol Biol. 2021;2290:215-228. doi: 10.1007/978-1-0716-1323-8_15.
Cloning proteins enables their production and characterization for further studies. This requires inserting the gene of the studied protein to be inserted in a vector, which then will be transformed to the host cell used as "factory." Consequently, the "biomass" of host cells will be produced using bioreactors. Here we describe the production of Rhizomucor miehei lipase (RML) by cloning the corresponding genes in the yeast Pichia pastoris. This enzyme is used as a biocatalyst for biofuel production. The successfully produced recombinant proteins are then purified using ion exchange chromatography.
克隆蛋白质可以使其生产和特性化，从而进行进一步的研究。这需要将研究的蛋白质的基因插入到载体中，然后将其转化为用作“工厂”的宿主细胞。因此，使用生物反应器生产宿主细胞的“生物质”。在这里，我们描述了通过在酵母毕赤酵母中克隆相应基因来生产根霉脂肪酶（RML）。该酶用作生物燃料生产的生物催化剂。然后使用离子交换色谱法对成功生产的重组蛋白进行纯化。
