论文部分内容阅读
目的建立一种基于蛋白重复序列及线性B细胞表位预测筛选间日疟原虫(Plasmodium vivax,P.vivax)特异性抗原肽的方法。方法以PlasmoDB数据为基础,建立间日疟原虫蛋白序列数据库。编写重复序列查找软件,逐一检索16 aa多肽在全部蛋白序列中的重复次数,统计出重复较高者进行连续B细胞表位预测,优选具有间日疟原虫特异性的肽进行合成,偶联钥孔戚血蓝蛋白(Keyhole limpet hemocyanin,KLH)后免疫小鼠,并检测抗体滴度。结果通过软件分析间日疟原虫全部5 432个蛋白序列中的16肽的重复次数,利用BcePred网站预测其中重复较高的1 000个序列,获得22个候选肽,经聚类分析和相似性比较,优选到5个潜在特异性肽,人工合成并偶联KLH后免疫小鼠诱生的抗体滴度均超过1:9 000。结论建立了一种基于蛋白重复序列联合线性B细胞表位预测筛选间日疟原虫特异性抗原肽的方法,得到的5个肽均能诱导小鼠产生高滴度抗体。
OBJECTIVE: To establish a method for screening Plasmodium vivax (P. vivax) specific antigen peptides based on protein repeat sequences and linear B cell epitopes. Methods Based on PlasmoDB data, a database of protein sequences of Plasmodium vivax was established. Sequencing software was designed to search the number of repetitions of the 16 aa polypeptide in all the protein sequences one by one. The repeated higher B-cell epitopes were predicted, and peptides with specificity of Plasmodium vivax were synthesized. Mice were immunized after immunization with Keyhole limpet hemocyanin (KLH) and antibody titers were measured. Results The software was used to analyze the number of repetitions of 16 peptides in all 5 432 protein sequences of Plasmodium vivax and 22 candidate peptides were predicted using the BcePred website, and 22 candidate peptides were obtained. By cluster analysis and comparability , Preferably up to 5 potential specific peptides. Immunization of mice after artificial synthesis and conjugation of KLH yielded antibody titers in excess of 1: 9,000. Conclusion A novel method based on protein sequence repeats and linear B cell epitopes screening for specific antigenic peptide of Plasmodium vivax was established. All the five peptides obtained could induce high titer antibody in mice.