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恶性疟原虫(P.f.)获取核酸并进行复制的机制是抗疟化疗的一个潜在中心环节。由于疟原虫不能从外界获得嘧啶,只能依靠自身合成,已确定在P.f.中有合成尿苷酸的酶类,但尚未纯化出来。作者已经分离了嘧啶合成中第一个酶——氨甲酰磷酸合成酶Ⅱ(CPSⅡ)的编码基因。已知CPS是一种极不稳定的酶,难以测到其活性。本文报告一种微量CPS检测方法。 在2%血细胞比容中培养FCQ-27和K1株恶性疟原虫,同步发育至成熟滋养体后,收获虫体并萃取蛋白。对于大量检测,取
The mechanism by which P. falciparum acquires nucleic acid and replicates is a potential central link to anti-malarial chemotherapy. Since P. falciparum can not obtain pyrimidine from the outside world and can only rely on its own synthesis, enzymes that synthesize uridine acid have been identified in P.f., but have not yet been purified. The authors have isolated the coding gene for the first enzyme, carbamylphosphate synthase II (CPS II), in pyrimidine synthesis. CPS is known to be a very labile enzyme and its activity is difficult to measure. This article reports a trace CPS test method. FCQ-27 and K1 strains of P. falciparum were cultured in 2% hematocrit and developed to mature trophozoites synchronously. The parasites were harvested and proteins extracted. For a large number of tests, take