Militello Kevin T, Wirth Dyann F
Department of Immunology and Infectious Diseases, Building I Room 705, Harvard School of Public Health, 665 Huntington Avenue, Boston, MA 02115, USA.
Parasitol Res. 2003 Jan;89(2):154-7. doi: 10.1007/s00436-002-0721-5. Epub 2002 Aug 23.
Transfection of Plasmodium falciparum has remained difficult and laborious due to a lack of suitable reporter genes and low transfection efficiency. Therefore, the luciferase gene of Renilla reniformis, a sensitive mammalian reporter gene, was evaluated as a reporter gene in this system. Our studies indicate that the R. reniformis luciferase gene can be expressed in P. falciparum and is easily detected by luminometry. P. falciparum extracts do not contain endogenous R. reniformis luciferase activity, which is essential for its use as a reporter gene in this organism. Moreover, both firefly and R. reniformis luciferase genes can be co-expressed in P. falciparum and their respective enzyme activities can be measured from the same sample. Thus, the R. reniformis luciferase gene can be used as an experimental reporter gene and/or used in conjunction with the firefly luciferase gene where one gene would be used to control transfection efficiency. The R. reniformis luciferase gene thus provides a valuable tool to facilitate transient transfection analysis in P. falciparum.
由于缺乏合适的报告基因以及转染效率较低，恶性疟原虫的转染一直困难且费力。因此，在该系统中评估了一种灵敏的哺乳动物报告基因——海肾萤光素酶基因，作为报告基因。我们的研究表明，海肾萤光素酶基因可在恶性疟原虫中表达，并且通过发光测定法易于检测。恶性疟原虫提取物不含有内源性海肾萤光素酶活性，这对于其在该生物体中用作报告基因至关重要。此外，萤火虫萤光素酶基因和海肾萤光素酶基因均可在恶性疟原虫中共表达，并且它们各自的酶活性可从同一样品中测量。因此，海肾萤光素酶基因可用作实验报告基因和/或与萤火虫萤光素酶基因联合使用，其中一个基因可用于控制转染效率。海肾萤光素酶基因因此为促进恶性疟原虫中的瞬时转染分析提供了一种有价值的工具。
