Hasenkamp Sandra, Merrick Catherine J, Horrocks Paul
Institute for Science and Technology in Medicine, Keele University, Staffordshire ST5 5BG, United Kingdom.
Mol Biochem Parasitol. 2013 Feb;187(2):117-20. doi: 10.1016/j.molbiopara.2013.01.001. Epub 2013 Jan 11.
Genetic modification of Plasmodium falciparum is a key molecular tool for the investigation of the biology and pathogenesis of this important human pathogen. The most effective means to introduce exogenous DNA into P. falciparum is via passive uptake following invasion into a DNA-loaded erythrocyte. Using bioluminescence as a tool to quantify transfection efficiency, parameters previously judged empirically to enhance transfection efficiency were subjected to a quantitative analysis. This report supports roles for fresh erythrocytes and growth medium supplemented with human serum in enhancing transfection efficiency. Critically, a proposed enhancement to transfection efficiency through continued feeding with DNA-loaded erythrocytes is not borne out in this study, and actually appears to be detrimental.
恶性疟原虫的基因改造是研究这种重要人类病原体生物学和发病机制的关键分子工具。将外源DNA导入恶性疟原虫的最有效方法是在其侵入加载了DNA的红细胞后通过被动摄取实现。利用生物发光作为量化转染效率的工具，对先前凭经验判断可提高转染效率的参数进行了定量分析。本报告支持新鲜红细胞和补充了人血清的生长培养基在提高转染效率方面所起的作用。至关重要的是，本研究并未证实通过持续喂食加载了DNA的红细胞来提高转染效率这一设想，实际上这似乎是有害的。
