Miller A J, Zhou J J
Biochemistry and Physiology Department, IACR-Rothamsted, Harpenden, UK.
Biochim Biophys Acta. 2000 May 1;1465(1-2):343-58. doi: 10.1016/s0005-2736(00)00148-6.
The Xenopus oocyte provides a powerful system for the expression and characterisation of plant membrane proteins. Many different types of plant membrane proteins have been expressed and characterised using this system. As there are already several general reviews on the methodology for oocyte expression of channel proteins, we have summarised the particular advantages and disadvantages of using the system for the characterisation of plant cotransporter proteins. As an example of how the system can be used to identify transporters, we describe evidence for a low affinity nitrate transporter in oocytes injected with poly(A) RNA extracted from nitrate-induced barley roots. Furthermore, we describe evidence that the expression of some transporters in oocytes can modify the properties of endogenous membrane proteins. We conclude that although care must be taken in the interpretation of results and in choosing appropriate controls for experiments, oocyte expression is an excellent tool which will have an important role in characterising plant membrane proteins.
非洲爪蟾卵母细胞为植物膜蛋白的表达和特性研究提供了一个强大的系统。许多不同类型的植物膜蛋白已通过该系统进行了表达和特性研究。由于已有多篇关于通道蛋白卵母细胞表达方法的综述,我们总结了使用该系统表征植物共转运蛋白的具体优缺点。作为该系统如何用于鉴定转运蛋白的一个例子,我们描述了在注射从硝酸盐诱导的大麦根中提取的聚腺苷酸RNA的卵母细胞中存在低亲和力硝酸盐转运蛋白的证据。此外,我们还描述了一些转运蛋白在卵母细胞中的表达可改变内源性膜蛋白特性的证据。我们得出结论,尽管在结果解释和实验合适对照的选择上必须谨慎,但卵母细胞表达是一个出色的工具,将在植物膜蛋白特性研究中发挥重要作用。