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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。
Apple stem pitting virus (ASPV) is a latent virus that seriously endangers the production of fruit trees, but lacks practical ASPV antiserum. Using different bioinformatics softwares, the antigen index of different regions of ASPV coat protein, α-helix, β-sheet, β-turn, irregular curly structure, hydrophilicity, flexibility and surface accessibility Surface probability. Based on the fact that the antigen epitopes are mainly distributed in the β-turn structure and the irregular curly region, the hydrophilicity and the surface accessibility parameters are higher. The comprehensive analysis predicts that the epitope of the ASPV coat protein epitope may be located at the N-terminal 6-20,100- Near the 114,400-414 amino acid residues. Two antisera (1468 and 1469) were finally obtained by synthesizing two polypeptides (CRGYEEGSRPNQRVLP, CTGGKIGPKPVLSIRK) and conjugating with carrier protein. The prepared antiserum can produce immunoreaction with prokaryotic expression product of ASPV coat protein gene. Application of this antiserum to the detection of apple samples, antiserum 1468 test results and RT-PCR test results, and the antiserum 1469 can only detect some samples.