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当前位置: 首页 > 文章 > 基于抗原表位策略的苹果茎痘病毒抗血清的制备 植物病理学报 2010,40 (4) 357-363
Position: Home > Articles > Production of antiserum of Apple stem pitting virus based on antigenic epitopes technique Acta Phytopathologica Sinica 2010,40 (4) 357-363

基于抗原表位策略的苹果茎痘病毒抗血清的制备

作  者：
李丽丽;杨洪一;董雅凤;高源
单  位：
沈阳农业大学园艺学院;中国农业科学院果树研究所;东北林业大学生命学院
关键词：
苹果茎痘病毒;表位;抗血清制备
摘  要：
苹果茎痘病毒(Apple stem pitting virus,ASPV)是一种严重危害果树生产的潜隐性病毒,但缺乏实用的ASPV抗血清。应用不同生物信息学软件对ASPV外壳蛋白不同区域的抗原指数、蛋白二级结构中α螺旋、β片层、β转角、无规则卷曲结构、亲水性、可塑性(Flexibility)和表面可及性(Surface probability)进行了分析。根据抗原表位主要分布在β转角结构和无规则卷曲区域、亲水性和表面可及性参数较高的特点,综合分析预测ASPV外壳蛋白抗原表位区可能位于N端6-20、100-114、400-414位氨基酸残基附近。通过合成2条多肽(CRGYEEGSRPNQRVLP、CTGGKIGPKPVLSIRK),与载体蛋白偶联后免疫动物,最后得到了2份抗血清(编号为1468和1469)。制备的抗血清可与ASPV外壳蛋白基因原核表达产物产生免疫反应。应用此抗血清检测苹果样品,抗血清1468检测结果与RT-PCR检测结果一致,而抗血清1469仅能检测到部分样品。
译  名：
Production of antiserum of Apple stem pitting virus based on antigenic epitopes technique
作  者：
LI Li-li1,YANG Hong-yi2,DONG Ya-feng3,GAO Yuan3(1College of Horticulture,Shen-yang Agricultural University,Shenyang 110161,China;2College of Life Sciences,Northeast Forestry University,Harbin 150040,China;3Research Institute of Pomology,Chinese Academy of Agricultural Science,Xingcheng 125100,China)
关键词：
Apple stem pitting virus(ASPV);epitope;antiserum
摘  要：
Apple stem pitting virus(ASPV)is one of the most important latent viruses infected fruit trees.The antiserum for effective detection of ASPV had not been produced.In the study,some characteristics of coat protein(CP),such as the antigenic index,α-helix,β-sheet,β-turn,coil structure,hydrophilicity,surface probability and flexibility were analyzed with the algorithms in some softwares.Based on the rules for location of the antigenic epitopes in the regions including β-turn and coil structure with high hydrophilicity and surface probability,the predicted epitopes were located in the region of amino acid position 6-20,100-114 and 400-414 respectively.Two linear synthetic peptides(CRGYEEGSRPNQRVLP and CTGGKIGPKPVLSIRK)were prepared and conjugated with carrier protein.At last,the antisera 1468 and 1469 were obtained by immunizing the rabbits.The antibody producted strong immunoreaction with the expression product of ASPV coat protein gene in Escherichia coli.The result of application of the antiserum 1468 in ELISA was the same as that of RT-PCR for detecting ASPV,but only partial positive samples could be detected with the antiserum 1469.

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