嗜肺军团菌momp原核重组质粒的构建和表达

The Construction and Expression of Recombinant Plasmid pET-momp of Legionella Pneumophila Major Outer Membrane Protein Gene

目的构建嗜肺军团菌momp原核重组质粒,并纯化重组蛋白。方法以嗜肺军团菌LP1型DNA为模板,PCR扩增得到momp基因,定向克隆至原核载体PET32a(+)中,经酶切及测序鉴定正确后,转化入大肠杆菌BL21中,用IPTG诱导并用SDS-PAGE电泳进行分析,其产物用亲和层析法进行纯化。结果扩增出831bp的momp基因,构建重组质粒pET-momp,诱导表达及纯化出50KD的蛋白。结论成功构建momp基因的原核表达载体并得到高效表达。

Objective To construct recombinant plasmid pET-momp of Legionella Pneumophila major outer membrane protein Gene and to purify the recombinant protein.Methods LP1 type of Legionella pneumophila DNA（a template momp gene） was amplified by PCR and cloned into prokaryotic vector PET32a（＋）.After restriction analysis and DNA sequencing,It was transformed into E.coli BL21,then it was induced with IPTG and analyzed by SDS-PAGE.Its products were purified by affinity chromatography.Results 831bp momp gene was amplified and,the recombinant plasmid pET-momp was constructed.50kD MOMP protein was successfully purified.Conclusion Momp gene was successfully constructed and highly expressed.