Wallach M, Cully D F, Haas L O, Trager W, Cross G A
Mol Biochem Parasitol. 1984 May;12(1):85-94. doi: 10.1016/0166-6851(84)90046-x.
The presence of histidine-rich protein (HRP) related genes and gene products in Plasmodium falciparum was demonstrated using a synthetic pentahistidine-encoding oligonucleotide and a cloned HRP cDNA probe prepared from the avian parasite P. lophurae. In Northern blotting experiments, two knobby clones of P. falciparum were found to contain a 3500 nucleotide RNA species that hybridized with the oligonucleotide and HRP cDNA probes. As this component had the expected size for an mRNA encoding an 80-90 kDa protein and was absent from two knobless clones of P. falciparum, we concluded that it represented a 'knob protein' mRNA. Using the restriction enzyme EcoRI, three identical cross-hydribizing HRP gene fragments were found in the DNA of both knobby and knobless clones of P. falciparum. These fragments differed in size from those present in P. lophurae. These results suggest that the absence of knob protein mRNA in knobless clones is not due to loss of the corresponding gene(s).
利用合成的编码五组氨酸的寡核苷酸以及从禽类疟原虫——绿头鸭疟原虫制备的克隆的富含组氨酸蛋白(HRP)cDNA探针,证实了恶性疟原虫中存在富含组氨酸蛋白相关基因及其基因产物。在Northern印迹实验中,发现恶性疟原虫的两个有突起克隆含有一种3500个核苷酸的RNA分子,它能与寡核苷酸和HRP cDNA探针杂交。由于该成分具有编码80 - 90 kDa蛋白质的mRNA预期大小,且在恶性疟原虫的两个无突起克隆中不存在,我们得出结论,它代表一种“突起蛋白”mRNA。利用限制性内切酶EcoRI,在恶性疟原虫有突起和无突起克隆的DNA中均发现了三个相同的交叉杂交HRP基因片段。这些片段的大小与绿头鸭疟原虫中的片段不同。这些结果表明,无突起克隆中缺乏突起蛋白mRNA并非由于相应基因的缺失。