Lee M G, Van der Ploeg L H
Division of Tropical Medicine, School of Public Health, Columbia University, New York, NY 10032.
Science. 1990 Dec 14;250(4987):1583-7. doi: 10.1126/science.2177225.
Development of methods for the manipulation of the genomes of parasitic protozoa will lead to enhanced understanding of parasite biology and host-parasite relationships. Efficient gene transfer and targeted integration by homologous recombination were achieved in the parasitic protozoan Trypanosoma brucei, the causative agent of sleeping sickness. An expression vector with the neomycin phosphotransferase gene (neo), under the control of a procyclic acidic repetitive protein (PARP) gene promoter, was targeted into an intergenic region in beta alpha-tubulin-gene tandem array. Sixteen copies of neo were found in a tandem array in one of the transfectants where the PARP promoter controlled alpha-amanitin-resistant transcription of neo, whereas transcription of tubulin genes remained alpha-amanitin-sensitive.
开发用于操纵寄生原生动物基因组的方法将有助于增强对寄生虫生物学以及宿主 - 寄生虫关系的理解。在昏睡病的病原体——寄生原生动物布氏锥虫中实现了高效的基因转移和通过同源重组进行的靶向整合。一个带有新霉素磷酸转移酶基因（neo）的表达载体，在一个前循环酸性重复蛋白（PARP）基因启动子的控制下，被靶向插入到β-微管蛋白 - 基因串联阵列中的一个基因间隔区。在其中一个转染子中发现了16个串联排列的neo拷贝，在该转染子中PARP启动子控制着neo对α-鹅膏蕈碱抗性的转录，而微管蛋白基因的转录对α-鹅膏蕈碱仍敏感。
