人成熟叠朊在大肠埃希菌中高效表达

Expression of Human Mature Prion-like Protein Doppel in Escherichia coli

以含有人叠朊编码基因Prnd的ORF的质粒HoPrnd-T为模板,PCR扩增出成熟叠朊编码基因片段homDpl,并将其插入原核表达载体pGEX-6P-1的EcoRⅠ和XhoⅠ位点之间,构建出重组表达质粒pGEX-homDpl。将pGEX-homDpl电转化到宿主菌株BL21(DE3)中,以1.0 mmol/LIPTG诱导表达,SDS-PAGE结果显示人成熟叠朊被大肠埃希菌高效表达。

HomDpl, the encoding gene of human mature prion-like protein Dopple, was amplified by PCR with the plasmid HoPrnd-T including the ORF of the human Prnd gene as a template. Then HomDpl was inserted into the EcoR Ⅰ - Xho Ⅰ site of pGEX-6P-1 vector to generate recombinant expression plasmid pGEX-homDpl. The pGEX-homDpl plasmid was transformed into host strain E. coli BL21 （DE3） through electroporation. 1.0 mmol/L IPTG was used to induce the recombinant E. coli to express homDpl protein. SDS-PAGE analysis demonstrated that homDpl protein was overexpressed in E. coli .