棉铃虫类肌钙蛋白多克隆抗体的制备与鉴定

doi:10.13560/j.cnki.biotech.bull.1985.2017-1115

摘要/Abstract

摘要： 利用蛋白免疫法制备鼠抗棉铃虫类肌钙蛋白（Helicoverpa armigera Calponin,HaCal）的多克隆抗体并进行初步应用。通过原核表达棉铃虫类肌钙蛋白,镍柱纯化获得目的蛋白,经足垫加皮下注射免疫小鼠,4次免疫后用ELISA检测法确定鼠抗HaCal蛋白多克隆抗体的效价。通过Western blot和免疫组化检测该多克隆抗体免疫特异性。蛋白免疫法制备的鼠抗HaCal多克隆抗体滴度高于1∶819 200。Western-blot和免疫组化显示该抗体能特异性识别棉铃虫体内的天然HaCal蛋白。利用蛋白免疫法制备获得了效价高且特异性强的鼠抗棉铃虫HaCal多克隆抗体,旨为进一步研究棉铃虫HaCal蛋白表达特性及其抗药性功能提供重要的实验工具。

关键词: 棉铃虫, 类肌钙蛋白, 多克隆抗体

Abstract: Polyclonal antibodies against calponin from Helicoverpa armigera（HaCal）were prepared by protein immunization and used for preliminary application. The HaCal was expressed by the prokaryotic expression system,purified by Ni-column,and then immunized the mice by footpad and subcutaneous injection. The titer of polyclonal antibody against HaCal protein was determined by ELISA after four immunizations. The immunological specificity of the polyclonal antibody was detected by Western blot and immunohistochemical staining. The titer of mouse anti-HaCal polyclonal antibody prepared by protein immunization was higher than 1：819 200. Western blot and immunohistochemical staining showed that the antibody specifically recognized the native HaCal protein in the H. armigera. In summary,the polyclonal antibody against HaCal with high titer and specificity was prepared by protein immunization,which provided an important experimental tool for further studying the expression characteristics of HaCal protein and clarifying how to invovle in drug resistance of H. armigera.

Key words: Helicoverpa armigera, calponin, polyclonal antibody

尼加提·迪里木拉提, 赵洁, 艾新宇, 刘小宁. 棉铃虫类肌钙蛋白多克隆抗体的制备与鉴定[J]. 生物技术通报, 2018, 34(7): 154-159.

Nijiati·DILIMULATI, ZHAO Jie, AI Xin-yu, LIU Xiao-ning. Preparation and Identification of Polyclonal Antibody Against Calponin from Helicoverpa armigera[J]. Biotechnology Bulletin, 2018, 34(7): 154-159.

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