Hitzeroth Inga I, van Zyl Albertha R
Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, Private Bag, Rondebosch, 7700, South Africa.
Methods Mol Biol. 2016;1404:581-595. doi: 10.1007/978-1-4939-3389-1_38.
Transient expression of viral proteins in plants is a novel alternative to other expression platforms. The viral proteins can be used as potential vaccines or in diagnostics. Nicotiana benthamiana leaves or whole plants are infiltrated with recombinant Agrobacterium that harbor the gene of interest. Protein expression in the plants is rapid and results are obtained within 2-7 days. Here we describe how to make electrocompetent Agrobacterium, how to transform Agrobacterium, how to infiltrate leaves or plants with the recombinant Agrobacterium, and lastly how to extract the protein for analysis by gel electrophoresis.
病毒蛋白在植物中的瞬时表达是一种有别于其他表达平台的新方法。病毒蛋白可用作潜在疫苗或用于诊断。将携带目的基因的重组农杆菌渗入本氏烟草叶片或整株植物中。植物中的蛋白表达迅速，2至7天内即可获得结果。在此，我们描述了如何制备电转化感受态农杆菌、如何转化农杆菌、如何用重组农杆菌渗入叶片或植物，以及最后如何提取蛋白用于凝胶电泳分析。
