TRANSFECTION OF PLASMODIUM-FALCIPARUM WITHIN HUMAN RED-BLOOD-CELLS
TRANSFECTION OF PLASMODIUM-FALCIPARUM WITHIN HUMAN RED-BLOOD-CELLS
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DOI：
10.1073/pnas.92.4.973
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发表时间：
1995-02-14
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;SIFRI, CD;WELLEMS, TE
恶性疟原虫疟疾寄生虫内的人红细胞（RBC）已成功地转染产生氯霉素乙酰转移酶（CAT）。使用寄生的RBC的电穿孔来引入质粒，所述质粒具有侧接恶性疟原虫hsp86、hrp3和hrp2基因的5'和3'非翻译序列的CAT编码DNA。这些侧翼序列是表达所必需的，因为它们的切除消除了转染寄生虫中的CAT活性。来自天然CAT编码DNA的转染信号与来自适应于恶性疟原虫主要密码子偏好的合成DNA序列的转染信号进行了很好的比较，证明了细菌序列的有效表达，尽管其使用了罕见的恶性疟原虫密码子。转染的环阶段的寄生虫产生CAT信号至少一样强的转染的红细胞阶段的寄生虫，即使环阶段被更多的红细胞胞质比红细胞包围。红细胞期恶性疟原虫的转染提高了我们对这一主要病原体进行遗传分析的能力。
Plasmodium falciparum malaria parasites within human red blood cells (RBCs) have been successfully transfected to produce chloramphenicol acetyltransferase (CAT). Electroporation of parasitized RBCs was used to introduce plasmids that have CAT-encoding DNA flanked by 5' and 3' untranslated sequences of the P. falciparum hsp86, hrp3, and hrp2 genes. These flanking sequences were required for expression as their excision abolished CAT activity in transfected parasites. Transfection signals from native CAT-encoding DNA compared well with those from a synthetic DNA sequence adapted to the P. falciparum major codon bias, demonstrating effective expression of the bacterial sequence despite its use of rare P. falciparum codons. Transfected ring-stage parasites produced CAT signals at least as strong as transfected schizont-stage parasites even though ring stages are surrounded by more RBC cytoplasm than schizonts. The transfection of erythrocyte-stage P. falciparum parasites advances our ability to pursue genetic analysis of this major pathogen.