小鼠脂联素受体-1的生物信息学分析及其抗体制备.pdfVIP

156 生理学报 Acta Physiologica Sinica, February 25, 2008, 60 (1): 156-160 实验技术 小鼠脂联素受体 -1 的生物信息学分析及其抗体制备 1 2 2 2 2,* 许言午 ，闫莉 ，刘慧 ，武烨 ，吴博威 1 2 山西医科大学 生物化学与分子生物学教研室； 生理学教研室，太原 030001 摘 要：本文旨在根据小鼠脂联素受体-1 (adiponectin receptor-1, AdipoR-1) 的分子结构，制备抗AdipoR-1 的多克隆抗体， 为AdipoR-1 的进一步研究创造条件。利用生物信息学分析预测小鼠AdipoR-1 的理化特性、跨膜结构和抗原表位，结合 AdipoR-1 和AdipoR-2 氨基酸序列相似性比对结果，设计出一段由16 个氨基酸组成的多肽，用此肽段免疫大鼠得到多克隆 抗体，利用ELISA、Western blot 等方法鉴定其特异性和滴度，并将该抗体用于正常及高胆固醇血症小鼠肌肉组织AdipoR-1 的表达检测。结果表明，此抗体效价较高、特异性强。以上结果提示，生物信息学分析能较好地预测小鼠AdipoR-1 的 抗原表位，并成功制备高特异性的多克隆抗体。 关键词：脂联素受体- 1 ；小鼠；抗体；生物信息学 中图分类号：R 9 3 2 Bioinformatics analysis of mouse adiponectin receptor-1 and its antibody preparation 1 2 2 2 2,* XU Yan-Wu , YAN Li , LIU Hui , WU Ye , WU Bo-Wei 1Department of Biochemistry and Molecular Biology; 2Department of Physiology, Shanxi Medical University, Taiyuan 030001, China Abstract: To establish a method for preparation of anti-mouse adiponectin receptor-1 (AdipoR-1) polyclonal antibody, the polypeptide antigen corresponding to AdipoR-1 was designed by bioinformatics analysis. The possible physicochemical property and trans- membrane structure were predicted by ExPASy and TMHMM, respectively. The antigen epitopes of mouse AdipoR-1 and its immunogenicity were analyzed by Antigenic Prediction and AntigenProfiler, respectively. According to the similarity analysis between AdipoR-1 and AdipoR-2 by Clustal W, a 16-amino acid polypeptide was designed as the antigen corresponding to AdipoR-1. To ensure the specificity of the polypeptide antigen, similarity search was run in the protein databases such as SWISS-PROT, PDB and Prosite databases. The polypeptide synthes