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| 恶性疟原虫EBA-175与血型糖蛋白A结合位点的鉴定 | |
| 引用本文： | 孙晓敏,郝文波,李明,罗仁,贾钰华.恶性疟原虫EBA-175与血型糖蛋白A结合位点的鉴定[J].南方医科大学学报,2007,27(11):1696-1698. |
| 作者姓名： | 孙晓敏 郝文波 李明 罗仁 贾钰华 |
| 作者单位： | 1. 南方医科大学,生物技术学院,510515;南方医科大学,中医药学院,510515 2. 南方医科大学,生物技术学院,510515 3. 南方医科大学,中医药学院,510515 |
| 摘 要： | 目的 探索EBA-175与GPA之间的结合信息,为疟疾短肽疫苗及拮抗药物的研制奠定基础.方法 以EBA-175重组蛋白为靶,采用亲和筛选法对噬菌体随机十二肽库进行3轮筛选,通过ELISA、竞争抑制试验、Dot-ELISA及Western blotting等方法鉴定获得的噬菌体短肽与EBA-175之间的结合特性.对阳性克隆进行DNA序列测定,推导其十二肽的氨基酸序列并与GPA氨基酸全序列进行了同源性比较.结果 经3轮亲和筛选后,结合噬菌体得到良好富集.从第3轮洗脱液铺制的琼脂板中随机挑取30个噬菌体单克隆,ELISA检测有27个为阳性,阳性率达90%.竞争性ELISA显示多数阳性噬菌体能竞争抑制EBA-175与其单抗结合.DNA及氨基酸序列分析表明24个噬菌体展示十二肽中共有序列IRR与GPA的114-116位氨基酸同源.结论 阳性噬菌体表达的短肽是EBA-175所识别的模拟表位,IRR几位氨基酸可能对EBA-175与GPA的结合起重要作用. |
| 关 键 词： | 恶性疟原虫 EBA-175 血型糖蛋白A 噬菌体随机肽库 |
| 文章编号： | 1673-4254（2007）11-1696-03 |
| 修稿时间： | 2007-05-21 |
Identification of the binding site on glycophorin A for Plasmodium falciparum EBA-175 |
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| SUN Xiao-min,HAO Wen-bo,LI Ming,LUO Ren,JIA Yu-hua.Identification of the binding site on glycophorin A for Plasmodium falciparum EBA-175[J].Journal of Southern Medical University,2007,27(11):1696-1698. | |
| Authors: | SUN Xiao-min HAO Wen-bo LI Ming LUO Ren JIA Yu-hua |
| Affiliation: | 1 School of Biotechnology, 2 School of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515, China |
| Abstract: | Objective To identify the binding site on glycophorin A(GPA) for EBA-175 to provide clue for developing short peptide vaccine and therapeutic agents against Plasmodium falciparum.Methods With the recombinant protein of EBA-175 as the target molecule,the mimetic peptides of GPA were screened from a 12-mer random peptide library.Three rounds of biopanning were carried out,and enzyme-linked immunosorbent assay(ELISA),competitive ELISA,Dot-ELISA and Western blotting used to evaluate the binding between the phage-borne peptides and EBA-175.The insert DNA sequences of positive clones were determined and their amino acid sequences deduced.Results Thirty clones from the third round were randomly selected,of which 27 were found positive by sandwich ELISA.Competitive ELISA proved that most of the phage-borne peptides could competitively inhibit the binding of antibody(EBA-175 Ab) with EBA-175.Analysis of DNA and amino acid sequences indicated that 24 positive phage clones contained the conservative sequence of IRR,which was highly homologous with the 114-116 amino acids of GPA.Conclusion These phage-displayed peptides can bind with EBA-175,and the amino acid sequence IRR might play an important role in the binding between EBA-175 and GPA. |
| Keywords: | EBA-175 Plasmodium falciparum glycophorin A phage-displayed random peptide library |
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