Barnes D A, Thompson J, Triglia T, Day K, Kemp D J
Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia.
Mol Biochem Parasitol. 1994 Jul;66(1):21-9. doi: 10.1016/0166-6851(94)90032-9.
Many lines of Plasmodium falciparum undergo a deletion of the right end of chromosome 9 during in vitro cultivation accompanied by loss of cytoadherence to melanoma cells. The deletion also results in loss of expression of PfEMP1, the putative cytoadherence ligand, suggesting that PfEMP1 or a regulatory gene controlling PfEMP1 expression is encoded in this region. Initially a library of short fragments highly enriched for the right arm of chromosome 9 was constructed in bacteriophage lambda. Clones from this library were obtained randomly by the polymerase chain reaction (PCR) technique, sequenced and used to screen a yeast artificial chromosome (YAC)-P. falciparum library by PCR so that the region could be cloned and physically mapped in detail. We have used probes from this region to demonstrate that clones derived from ITG2 have undergone a deletion of intermediate length on chromosome 9. This could explain the unusual stability of cytoadherence in these clones.
许多恶性疟原虫株在体外培养过程中会发生9号染色体右端的缺失,同时伴随着对黑色素瘤细胞细胞粘附性的丧失。这种缺失还导致假定的细胞粘附配体PfEMP1表达缺失,这表明PfEMP1或控制PfEMP1表达的调控基因在该区域编码。最初,构建了一个在噬菌体λ中高度富集9号染色体右臂的短片段文库。通过聚合酶链反应(PCR)技术从该文库中随机获得克隆,进行测序,并用于通过PCR筛选酵母人工染色体(YAC)-恶性疟原虫文库,以便对该区域进行克隆和详细的物理图谱绘制。我们使用来自该区域的探针证明,源自ITG2的克隆在9号染色体上发生了中等长度的缺失。这可以解释这些克隆中细胞粘附的异常稳定性。