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Prokaryotic Expression and Purification of LscW Protein of Lawsonia intracellularis for biomedical engineering

机译:细胞内劳森菌LscW蛋白在生物医学工程中的原核表达和纯化

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In order to study the LscW of Lawsonia intracellularis, the prokaryotic expression vector of LscW protein was constructed. Two primers were designed in this experiment. The 1197 bp lscW gene was amplified from L. intracellularis B3903 strain by PCR and inserted into pCold expression vector to construct recombinant plasmid pCold/lscW, which was transformed into BL21 competent cells. The positive clones were induced by IPTG, purified by protein purification column, and confirmed by SDS-PAGE gel electrophoresis. The LscW was expressed in the supernatant and purified to obtain a single target protein band.
机译:为了研究胞内劳森菌的LscW,构建了LscW蛋白的原核表达载体。在该实验中设计了两种引物。通过PCR从胞内劳森氏菌B3903菌株中扩增出1197 bp的lscW基因,并将其插入pCold表达载体中,构建重组质粒pCold / lscW,并将其转化为BL21感受态细胞。阳性克隆经IPTG诱导,经蛋白纯化柱纯化,并经SDS-PAGE凝胶电泳确认。 LscW在上清液中表达并纯化以获得单个靶蛋白带。

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