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目的 分析人类腺病毒 (HAdV)六邻体保守区氨基酸序列 ,合成保守区多肽并制备抗多肽抗体 ,以确定型间共同的特异性表位。方法 用ClustalX、Antheprot等软件 ,对HAdV六邻体进行分析 ,确定保守区并分析其抗原性。合成抗原性多肽 ,免疫家兔制备抗肽多克隆抗体。用间接免疫荧光法和Westernblot法 ,检测所制备的抗肽抗体与病毒抗原的反应性。结果 根据HAdV 2六邻体的保守区合成 9种肽段 ,分成 3组免疫家兔 ,共获得 3组抗肽抗体。所有的抗肽抗体在 1∶10 0 0稀释度时 ,与 3个型别的HAdV感染细胞裂解物中相对分子质量 (Mr)约为 116 0 0 0的六邻体亚单位具有特异性反应。抗肽抗体对HAdV感染细胞有特征性荧光染色。 3组抗肽抗体中 ,有 7个抗体与 3个型别的HAdV感染细胞均具有阳性染色反应。结论 合成的保守区多肽能够诱导抗肽抗体产生 ,诱导产生的抗体与HAdV 3、 4、 7具有免疫反应性。通过分析预测抗原表位并制备抗肽抗体是可行的
OBJECTIVE: To analyze the amino acid sequence of the conserved region of human adenovirus (HAdV) hexon, to synthesize the polypeptide of conserved region and to prepare the anti-peptide antibody to identify the common specific epitopes. Methods The HAdV hexon was analyzed by software such as ClustalX and Antheprot, the conserved region was identified and its antigenicity was analyzed. Antigenic peptide was synthesized and immunized rabbits to prepare anti - peptide polyclonal antibody. Indirect immunofluorescence and Western blotting were used to detect the reactivity of the prepared anti-peptide antibody with virus antigen. Results Nine kinds of peptides were synthesized according to the conserved region of HAdV 2 hexon and divided into three groups of immunized rabbits. Three groups of anti-peptide antibodies were obtained. All of the anti-peptide antibodies reacted specifically with the hexon subunits of about 116 000 in the lysate of 3 types of HAdV infected cells at a dilution of 1: 100. Anti-peptide antibodies have characteristic fluorescent staining of HAdV-infected cells. Among the three groups of anti-peptide antibodies, seven antibodies had positive staining reaction with three types of HAdV infected cells. Conclusion The synthetic conserved region peptides can induce the production of anti-peptide antibodies. The induced antibodies are immunoreactive with HAdV 3, 4, 7. It is feasible to analyze the predicted epitopes and prepare anti-peptide antibodies