Perdomo J Alejandro, Sales Cristina R G, Carmo-Silva Elizabete
Plant Science Department, Rothamsted Research, Harpenden, UK.
Lancaster Environment Centre, Lancaster University, Lancaster, UK.
Methods Mol Biol. 2018;1770:215-227. doi: 10.1007/978-1-4939-7786-4_12.
In this chapter, we describe a method to extract and quantify photosynthetic enzymes using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. The method is particularly suitable for characterizing altered protein amounts in leaves of plants produced from genetic engineering or gene-editing approaches. We focus on RuBisCO and RuBisCO activase, a molecular chaperone required to sustain the activity of RuBisCO and CO fixation, yet the method can be easily adapted to investigate other leaf proteins of interest.
在本章中，我们描述了一种使用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和免疫印迹法来提取和定量光合酶的方法。该方法特别适用于表征通过基因工程或基因编辑方法产生的植物叶片中蛋白质含量的变化。我们重点关注核酮糖-1,5-二磷酸羧化酶/加氧酶（RuBisCO）和RuBisCO活化酶，RuBisCO活化酶是维持RuBisCO活性和二氧化碳固定所需的分子伴侣，但该方法可以很容易地适用于研究其他感兴趣的叶片蛋白质。
