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蔡兴雁,李静惠,施文钧,等.日本血吸虫虫卵特异性蛋白的基因克隆及多态性分析[J].同济大学学报（医学版）,2010,31(1):17-22,31. [点击复制]
- CAI Xing-yan,LI Jing-huiz,SHI Wen-jun,et al.Molecular cloning and polymorphism analysis of egg specific genes of Schistosoma japonicum[J].同济大学学报（医学版）,2010,31(1):17-22,31. [点击复制]
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| 日本血吸虫虫卵特异性蛋白的基因克隆及多态性分析 |
| 蔡兴雁,李静惠,施文钧,李军,庄文佳,陈小平 |
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| (同济大学医学院免疫学教研室,上海,200092;同济大学医学院,上海,200092;同济大学医学院微生物教研室,上海,200092;同济大学医学院寄生虫教研室,上海,200092) |
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| 摘要: |
| 目的日本血吸虫虫卵中可能参与免疫调节的分子的克隆及其多态性分析。方法在NCBI数据库及国家人类基因组南方研究中心（Chinese National HumanGenome Center，CHGC）日本血吸虫基因数据库中筛选出两个类 IPSE（IL-4-inducing principle of S. mansoni eggs）基因（AY812212和AY223149）、一个类蛇毒过敏样蛋白基因（AY812797）和-个类亲环素蛋白基因（AY814078）。用RT—PCR检测表达谱。并从日本血吸虫虫卵cDNA文库中克隆这四个基因。定向克隆入质粒pET28-a，经双酶切鉴定筛选得重组的阳性克隆后测序。所得序列应用生物信息学分析软件VectorNTISuite7．0、Mega4．0和在线软件Rankpep进行初步分析。结果成功构建了4个重组质粒pET28a-223149，pET28a-812212，pET28a-812797和pET28a-814078，并测定和分析了这4个基因。结论日本血吸虫虫卵基因存在基因多态性，可能与免疫逃避有关。 |
| 关键词: 日本血吸虫 虫卵特异性蛋白 克隆 基因多态性 |
| DOI：10.3969/j.issn1008-0392.2010.01.004 |
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| 基金项目:国家"九七三"重点基础研究发展计划资助项目 |
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| Molecular cloning and polymorphism analysis of egg specific genes of Schistosoma japonicum |
| CAI Xing-yan,LI Jing-huiz,SHI Wen-jun,LI Jun,ZHUANG Wen-jia,CHEN Xiao-ping |
| (Dept. of Immunology, Tongji University School of Medicine, Shanghai 200092, China;;Dept. of Microbiology, Tongji University School of Medicine, Shanghai 200092, China ;;Dept. of Parasitology, Tongji University School of Medicine, Shanghai 200092, China) |
| Abstract: |
| Objective To obtain molecules that possibly involved in immunoregulation from the egg of Schistosoma japonicum and their polymorphism analysis. Methods Four genes were re- trieved from the NCBI database and the database of S. japonicum delivered by Chinese National Hu- man Genome Center at Shanghai. These four genes included AY812212 and AY223149 which were similar to IPSE （IL-4-inducing principle of S. mansoni eggs） in Schistosoma masoni; AY812797 with predicted sequence similar to venom allergen-like protein, and AY814078 with predicted sequence similar to cyclophilin. The target genes were cloned into a prokaryotic expression vector pET28-a from S. japonicum cDNA library. The positive recombinant colonies were screened and identified by endonuclease digestion and then sequenced. Vector NTI Suite 7.0, Mega 4.0 and online software Rankpep were used for sequence analysis. Results The recombinant plasmids pET28a-223149, pET28a-812212, pET28a-812797 and pET28a-814078 were succesfully constructed, sequenced and analyzed. Conclusion The results suggests that these four genes express mainly during S. japonicum egg stage and demonstrate that genetic polymorphism may possibly involve in immune evasion. |
| Key words: Schistosoma japonicum egg specific proteins cloning polymorphism |