Kelleher M, Tomley F M
Division of Molecular Biology, The Institute for Animal Health, Compton, Berkshire, UK.
Mol Biochem Parasitol. 1998 Nov 30;97(1-2):21-31. doi: 10.1016/s0166-6851(98)00128-5.
A transient transfection system has been developed for a member of the Apicomplexa, Eimeria tenella, using beta-galactosidase (betagal) from Escherichia coli as the reporter enzyme. Successfully expressed constructs contained sequences of the E. tenella microneme gene Etmic-1 fused to the coding region of lacZ. Transfectants expressing betagal were able to invade host cells and proceed through part of the life-cycle, forming schizonts from which merozoites were released. This indicated that transfectants could differentiate at least to first generation schizonts. However, this differentiation was delayed compared with unelectroporated sporozoites by approximately 15 h. Some merozoites arising from transfected sporozoites also expressed betagal. These results are encouraging for the development of a stable transfection system for E. tenella, using betagal as a reporter enzyme.
已开发出一种针对顶复门成员柔嫩艾美耳球虫的瞬时转染系统,该系统使用来自大肠杆菌的β-半乳糖苷酶(βgal)作为报告酶。成功表达的构建体包含与lacZ编码区融合的柔嫩艾美耳球虫微小体基因Etmic-1的序列。表达βgal的转染子能够侵入宿主细胞并经历部分生命周期,形成裂殖体,从中释放出裂殖子。这表明转染子至少可以分化为第一代裂殖体。然而,与未电穿孔的子孢子相比,这种分化延迟了约15小时。转染子孢子产生的一些裂殖子也表达βgal。这些结果对于开发以βgal作为报告酶的柔嫩艾美耳球虫稳定转染系统来说是令人鼓舞的。