豇豆胰蛋白酶抑制剂单克隆抗体的制备及初步应用

Preparation and application of monoclonal antibodies against cowpea trypsin inhibitor

目的 检测转豇豆胰蛋白酶抑制剂 (CpTI)基因水稻叶片中CpTI蛋白 ,来对转CpTI基因作物的检测建立初步的方法。方法 目前国际上主要通过基因检测的方法 ,而基因的检测可能存在假阳性的结果 ,而且基因的导入并不一定代表功能蛋白的表达 ,表达含量也为未可知。因此 ,通过检测转基因蛋白含量的多少是最直接和明确的方法。通过体外基因重组技术制备的CpTI蛋白来免疫动物得到单克隆抗体 ,就可通过westernblotting法检测转基因作物中的CpTI。结果 得到三株抗CpTI的单克隆抗体 ,利用这三种单克隆抗体的混合物对转CpTI蛋白的水稻叶片进行初步检测 ,得到了较为满意的结果。

Objective To detect cowpea trypsin inhibitor(CpTI) in transgenic rice leaves and build a possible method to assay transgenic plant with CpTI. Methods Nowadays gene assay is the most useful method to detect transgenic plants. But false positive may be got in gene assay and effective protein may not be produced. So assay of foreign protein is the most direct and conformable method. Monoclonal antibodies were got through traditional method of preparation. Results Three monoclonal antibodies were got. Detection of CpTI in transgenic rice leaves with these three antibodies mixes through Western blotting was conducted and the result was satisfying. Conclusion The method of using the mixes of three antibodies to detect the CpTI in plants through Western blotting was satisfying.