恶性疟原虫海南株翻译控制肿瘤蛋白(TCTP)基因在M15中的表达

EXPRESSING OF THE TRANSLATIONALLY CONTROLLED TUMOR PROTEIN GENE OF PLASMODIUM FALCIPARUM STRAIN HAINAN IN E. COLI M15

获得翻译控制肿瘤蛋白(TCTP)基因后与表达载体pQE-30重组,然后转化大肠杆菌M15,经IPTG诱导后表达TCTP融合蛋白,薄层扫描结果显示目的蛋白占菌体总蛋白的23.4%,目的蛋白中可溶性蛋白所占比例高于非溶性蛋白。Ni-NTA纯化后C端测序,结果C端13个氨基酸序列与由GenBank国际基因库中恶性疟原虫TCTP基因推导出的C端13个氨基酸序列及组成完全一致。结果表明成功地在M15中表达了恶性疟原虫海南株TCTP基因,为进一步研究提供了实验材料。

The translationally controlled tumor protein （TCTP） gene of Plasmodiumfalciparum recombined with the pQE-30 vector was transformed into the E. coli M15 and induced with IPTG to express TCTP. SDS-PAGE flying spot scanning results showed that target TCTP is 23.4% of total protein of E. coli M15, and soluble protein appeared more than insoluble one in SDS-PAGE gel. And then, the soluble target protein was purified with Ni-NTA column, and its C-terminal was sequenced with mass spectrum analysis. 13 amino acids of C-terminal is identical to the Plasmodiumfalciparum TCTP when blasted in GenBank.