| 大鼠感染血清免疫筛选弓形虫速殖子cDNA文库 |
| |
| 引用本文： | 蒋立平,吴翔,蔡力汀,王丹静,舒衡平.大鼠感染血清免疫筛选弓形虫速殖子cDNA文库[J].中国寄生虫学与寄生虫病杂志,2003,21(2):87-89. |
| |
| 作者姓名： | 蒋立平 吴翔 蔡力汀 王丹静 舒衡平 |
| |
| 作者单位： | 中南大学湘雅医学院寄生虫学教研室,长沙,410078 |
| |
| 摘 要： | 目的为弓形虫病疫苗的研制提供新的抗原分子。方法用弓形虫RH株速殖子感染大鼠,分离其血清作为探针筛选弓形虫cDNA文库,对阳性克隆的插入片段分别进行PCR扩增及DNA序列测定。结果从cDNA文库4×105~5个噬菌斑中筛选出13个阳性克隆,其插入片段大小分别为0.45～2.4kb。对L1、L2、L4和L5四个克隆进行测序,将所得序列查询基因库,结果,克隆L2与弓形虫P24主要抗原基因序列相同,L4与蔗糖丙酮酸磷酸激酶具有同源性,L1无任何相匹配的序列,为未曾报告过的新基因(GenBank登录号为AY180109),命名为T.g-R1。T.g-R1编码134个氨基酸的非跨膜蛋白。PROSCAN分析显示T.g-R1含有2个蛋白激酶C磷酸化位点,2个酪蛋白激酶Ⅱ磷酸化位点,1个肉豆酸酰化位点,1个微体细胞C端靶信号。L5为一小片段,无完整编码读框。结论阳性克隆的筛选和鉴定为抗弓形虫病疫苗的研制提供又一途径。
|
| 关 键 词： | 弓形虫 cDNA文库 免疫筛选 大鼠 |
| 文章编号： | 1000-7423(2003)-02-0087-03 |
| 修稿时间： | 2002年12月10 |
Immunological Screening of Toxoplasma Tachyzoite cDNA Expression Libraries with Serum from Infected Rats |
| |
| JIANG Li-ping,WU Xiang,CAI Li-ting,WANG Dan-jing,SHU Heng-ping.Immunological Screening of Toxoplasma Tachyzoite cDNA Expression Libraries with Serum from Infected Rats[J].Chinese Journal of Parasitology and Parasitic Diseases,2003,21(2):87-89. |
| |
| Authors: | JIANG Li-ping WU Xiang CAI Li-ting WANG Dan-jing SHU Heng-ping |
| |
| Affiliation: | Department of Parasitology, Xiangya School of Medicine, Central South University, Changsha 410078. |
| |
| Abstract: | OBJECTIVE: To screen and identify the potential candidates for the development of toxoplasmosis vaccine. METHODS: Rats were infected with Toxoplasma gondii (T. gondii) RH strain and their sera were used as a probe to screen T. gondii tachyzoite cDNA expression libraries. The positive clones were analyzed by PCR amplification and DNA sequencing. RESULTS: Thirteen positive clones were obtained from about 4 x 10(5) phage plagues after three rounds of screening. The size of the inserts ranged from 0.45 kb to 2.4 kb. A BLAST search of all available sequence databases using the partial sequences from four positive clones (L1, L2, L4, L5) showed that the sequence of L2 clone was identical with T. gondii P24 major antigen gene (TgP24). Clone L4 had a high homology with Saccharum officinarum pyruvate orthophosphate dikinase. There is no significant hit of any sequences to clone L1, suggesting that L1 could be a novel gene(GenBank accession number AY180109), named T.g-R1, which encodes a non-transmembrane protein with 134 amino acid open reading frame. PROSCAN analysis of the T.g-R1 amino acid sequence showed that this gene product contains two protein kinase C phosphorylation site, two casein kinase II phosphorylation site, one N-myristoylation site and one microbodies C-terminal targeting signal. Clone L5 was a small partial fragment. CONCLUSION: The identification of positive clones provides a possible way for the development of toxoplasmosis vaccine. |
| |
| Keywords: | Toxoplasma gondii cDNA library immunoscreening rat |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
| 点击此处可从《中国寄生虫学与寄生虫病杂志》浏览原始摘要信息 |
|
点击此处可从《中国寄生虫学与寄生虫病杂志》下载全文 |
|
