Cheesman S, McAleese S, Goman M, Johnson D, Horrocks P, Ridley R G, Kilbey B J
Institute of Cell and Molecular Biology, University of Edinburgh, UK.
Nucleic Acids Res. 1994 Jul 11;22(13):2547-51. doi: 10.1093/nar/22.13.2547.
The gene for topoisomerase II has been isolated from genomic libraries of strain K1 of the human malarial parasite, Plasmodium falciparum. The sequence reveals an open reading frame of 4194 nucleotides which predicts a polypeptide of 1398 amino acids. There are apparently no introns. The sequence is present as a single copy which has an identity of 47.4% and a similarity of 65.4% with its human homologue. Sequences conserved in topoisomerase II from other species are present in Pftopoisomerase II but in addition it has two adjacent asparagine-rich insertions which are unique to it. We have also detected asparagine-rich regions in the gene for PfDNA polymerase alpha. The gene for Pftopoisomerase II has been localised to chromosome 14 and northern analysis reveals a transcript of 5.8 kb. Two independent antisera raised in mice against glutathione-S-transferase fusion proteins containing the amino terminal portion of the malarial protein detect a weak band on western blots at about 160kDa, the expected size of the protein. Use of the same antisera for immunofluorescence analysis suggests that the protein is present at all stages of intraerythrocytic growth of the parasite.
已从人类疟原虫恶性疟原虫K1株的基因组文库中分离出拓扑异构酶II基因。该序列揭示了一个4194个核苷酸的开放阅读框,预测其编码一个由1398个氨基酸组成的多肽。显然没有内含子。该序列以单拷贝形式存在,与其人类同源物的同一性为47.4%,相似性为65.4%。在其他物种的拓扑异构酶II中保守的序列也存在于恶性疟原虫拓扑异构酶II中,但除此之外,它还有两个相邻的富含天冬酰胺的插入序列,这是它所特有的。我们还在恶性疟原虫DNA聚合酶α基因中检测到了富含天冬酰胺的区域。恶性疟原虫拓扑异构酶II基因已定位到14号染色体,Northern分析显示有一个5.8kb的转录本。用含有疟原虫蛋白氨基末端部分的谷胱甘肽-S-转移酶融合蛋白在小鼠中制备的两种独立抗血清,在Western印迹上检测到一条约160kDa的弱带,这是该蛋白的预期大小。使用相同的抗血清进行免疫荧光分析表明,该蛋白在疟原虫红细胞内生长的所有阶段均存在。