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植物转脂蛋白(LTP)是一类广泛存在于高等植物中的空间结构高度保守的碱性小分子蛋白,其确切功能和调节机制至今仍不清楚.本室从白菜中分离的钙调素结合蛋白10(CaMBP10),经序列分析被鉴定为植物转脂蛋白家族成员.近期研究结果表明,CaMBP10参与了植物的生物与非生物胁迫反应.为了深入探讨CaMBP10的抗性机制,确定植物中与其相互作用的蛋白质,本文拟建立胶体金标记CaMBP10的方法,通过凝胶覆盖分析,检测植物样品中的CaMBP10结合蛋白.为此,对标记反应的最适条件进行了优化,确定最佳条件为:交联剂戊二醛用量为0.034%,交联反应pH值为7.0,交联反应时间为40 min,胶体金颗粒度为10 nm,胶体金溶液的pH为7.0.本文确定建立了植物样品中CaMBP10结合蛋白的分析与鉴定方法.
Plant lipoprotein (LTP) is a kind of small basic protein which is widely conserved in higher plants and highly conserved in space structure. Its exact function and regulatory mechanism are still unclear. Calmodulin binding Protein 10 (CaMBP10) was identified as a member of the plant lipoprotein family by sequence analysis.Recent results show that CaMBP10 is involved in plant biotic and abiotic stress responses.In order to further explore the mechanism of CaMBP10 resistance, In this paper, the method of colloidal gold labeling of CaMBP10 was established, and the CaMBP10-binding protein in plant samples was tested by gel coverage analysis.Therefore, the optimal conditions for labeling reaction were optimized, and the optimal conditions were as follows: The dosage of glutaraldehyde was 0.034%, the pH value of cross-linking reaction was 7.0, the cross-linking reaction time was 40 min, the colloidal gold particle size was 10 nm and the pH of colloidal gold solution was 7.0. Binding protein analysis and identification methods.