小麦Rubisco抗体的制备和旗叶中Rubisco含量的ELISA测定

Rubisco(核酮糖-1,5-二磷酸羧化酶/加氧酶)为光合生物中的关键性酶。以小麦(Triticum aestivum L. cv.)叶片为材料，在(NH4)2SO4盐析和DEAE-Sepharose F F层析纯化的基础上添加了Sephacryl S 300分子筛，进一步纯化了Rubisco。 将纯化的Rubisco免疫大白兔制备了Rubisco的多克隆抗体， 经Western blot检测，Rubisco抗体与Rubisco呈免疫反应特异性。并运用Rubisco抗体，采用ELISA方法直接测定小麦旗叶生长发育过程中Rubisco含量的变化，结果表明：不同生长阶段，旗叶的Rubisco含量与光合速率大小呈正相关。该方法比免疫沉淀法更为快捷、准确。

Preparation of Rubisco Antibody and Determination of Rubisco Content of Flag Leaves in Wheat by ELISA

The ribulose 1, 5-bisphosphate carboxylase/oxygenase (Rubisco) is crucial to the overall modulation of photosynthesis. Rubisco was purified by (NH4)2SO4, DEAE-Sepharose F F and Sephacryl S 300 from wheat (Triticum aestivum L. cv.) leaves. The rabbit polyclonal antibody was prepared by vaccinating rabbit with purified Rubisco. The Rubisco antibody was showed specificity to Rubisco by Western blot. And using Rubisco antibody,contents variation of Rubisco in flag leaves was determined by ELISA. The result showed that Rubisco contents were positive relative with photosynthetic rates during development stages of flag leaves. This method is quicker and more accurate than that of immune precipitation.