Transformation of Plasmodium falciparum malaria parasites by homologous integration of plasmids that confer resistance to pyrimethamine
Transformation of Plasmodium falciparum malaria parasites by homologous integration of plasmids that confer resistance to pyrimethamine
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DOI：
10.1073/pnas.93.3.1130
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发表时间：
1996-02-06
影响因子：
11.1
通讯作者：
Wellems, TE
中科院分区：
文献类型：
--
作者：
Wu, YM;Kirkman, LA;Wellems, TE
用含有恶性疟原虫、恶性疟原虫或弓形虫二氢叶酸还原酶-胸腺酸合成酶(dhfr-ts)编码序列的质粒转化恶性疟原虫，在乙胺胺压力下，获得转化的疟原虫，这些转化的寄生虫在几周内保持未重排的上染色体，这些寄生虫群体在2-3个月后被将导入的DNA整合到核染色体中的寄生虫取代，根据转化所用的特定结构，在作为基因控制区的P、恶性疟原虫Dhfr基因座(染色体4)或hrp3和hrp2序列(分别为染色体13和8)中检测到同源整合。通过同源整合的转化为靶向基因改变和敲除奠定了基础，这将促进对恶性疟原虫的了解。
Plasmodium falciparum malaria parasites were transformed with plasmids containing P, falciparum or Toxoplasma gondii dihydrofolate reductase-thymidylate synthase (dhfr-ts) coding sequences that confer resistance to pyrimethamine, Under pyrimethamine pressure, transformed parasites were obtained that maintained the transfected plasmids as unrearranged episomes for several weeks, These parasite populations were replaced after 2 to 3 months by parasites that had incorporated the transfected DNA into nuclear chromosomes, Depending upon the particular construct used for transformation, homologous integration was detected in the P, falciparum dhfr-ts locus (chromosome 4) or in hrp3 and hrp2 sequences that were used in the plasmid constructs as gene control regions (chromosomes 13 and 8, respectively), Transformation by homologous integration sets the stage for targeted gene alterations and knock-outs that will advance understanding of P, falciparum.