耿超;牟安丽;刘志强;李向东;周涛;

• 1:山东农业大学植物保护学院植物病理学系
• 2:中国农业大学植病系

摘要(Abstract)：

【目的】为了制备苹果褪绿叶斑病毒(Apple chlorotic leaf spot virus,ACLSV)的抗血清,【方法】将ACLSV外壳蛋白(Coat protein,CP)基因克隆到原核表达载体pEHISTEV和pEHISEGFPTEV,转化大肠杆菌Rosetta,切胶回收诱导表达的CP融合蛋白免疫新西兰长耳兔,制备ACLSV的抗血清。【结果】利用原核表达载体pEHISTEV和pE-HISEGFPTEV在Rosetta中分别表达出了分子质量为25 ku和55 ku的ACLSV CP融合蛋白。切胶回收25 ku的融合蛋白免疫新西兰长耳兔。经ELISA测定,所制备的抗血清效价为1∶1 024,Western blot及组织印迹分析表明抗血清能与ACLSV CP发生特异性的免疫反应。【结论】该血清特异性强,效价高,可用于苹果叶片、果实和枝条等样品中A-CLSV的检测。

关键词(KeyWords)： 苹果褪绿叶斑病毒;外壳蛋白基因;原核表达;抗血清;组织印迹分析

Abstract:

Keywords:

基金项目(Foundation): 山东省自然科学基金(ZR2011CM019)

作者(Authors): 耿超;牟安丽;刘志强;李向东;周涛;

DOI: 10.13925/j.cnki.gsxb.2012.05.001

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