伯氏疟原虫类巨噬细胞移动抑制因子在红内期的表达

目的制备小鼠抗伯氏疟原虫类巨噬细胞移动抑制因子(PbMIF)的抗体,并检测PbMIF在伯氏疟原虫红内期的表达。方法用纯化的PbMIF蛋白免疫BABL/c小鼠,制备小鼠抗PbMIF的抗血清。用酶联免疫吸附试验(ELISA)检测抗体的效价,用免疫印迹(Western blotting)-二氨基联苯胺(DAB)底物显色法检测抗体的特异性。采用免疫印迹增强化学发光(ECL)的方法,用上述制备的抗血清检测PbMIF在红内期伯氏疟原虫的表达情况。结果ELISA测得小鼠抗PbMIF抗体的效价>1∶106,Western blotting显示该抗体可与原核表达的PbMIF蛋白特异性结合。ECL结果表明感染的红细胞中存在PbMIF。结论成功制备了小鼠抗PbMIF的抗体。红内期伯氏疟原虫表达PbMIF。

Expression of homologue of macrophage migration inhibitory factor from Plasmodium berghei in intra-erythrocyte stage

To determine the expression of homologue of macrophage miration inhibitory factor from Plasmodium berghei(PbMIF) in intra-erythrocyte stage,mouse antiserum against PbMIF was prepared,the purified PbMIF protein was used to immunize BALB/c mice to obtain anti-PbMIF serum.The titre of the polyclonal antibodies was determined by ELISA,and the specificity was assayed by Western blotting developed with chromogenic substrate 3,3'-diaminobenzidine(DAB).The expression of PbMIF in the intra-erythrocyte stage of P.berghei infection was detected with this antiserum by Western-blotting-enhanced chemiluminescence(ECL).The experimental results demonstrated that the PbMIF-specific antiserum showed high specificity and high titer of antibody activity.The PbMIF protein could be detected in the infected erythrocytes.