中国媒介生物学及控制杂志 ›› 2011, Vol. 22 ›› Issue (1): 44-48,51.

• 论著 • 上一篇    下一篇

日本血吸虫成虫吡喹酮用药前后差异表达序列的筛选

周水茂, 江元山, 陈智, 熊燕, 田俊华, 周敦金, 徐明星, 孙东光   

  1. 武汉市疾病预防控制中心微生物检验科,湖北 武汉 430015
  • 收稿日期:2010-09-28 出版日期:2011-02-20 发布日期:2011-02-20
  • 通讯作者: 江元山, Email: jiangshanis@yahoo.com.cn
  • 作者简介:周水茂(1967-),男,副主任技师,主要从事血吸虫病控制与防治研究。Email: 867494784@qq.com

Screening of differential expression of expressed sequence tags(ESTs) in adult Schistosoma japonicum treated with praziquantel

ZHOU Shui-mao, JIANG Yuan-shan, CHEN Zhi, XIONG Yan, TIAN Jun-hua, ZHOU Dun-jin, XU Ming-xing, SUN Dong-guang   

  1. Wuhan Center for Disease Control and Prevention, Wuhan 430015, Hubei Province, China
  • Received:2010-09-28 Online:2011-02-20 Published:2011-02-20

摘要:

目的 应用抑制性消减杂交(SSH)技术构建吡喹酮治疗日本血吸虫感染新西兰大白兔前后肝脏内差减cDNA文库,为筛选吡喹酮治疗过程中日本血吸虫体内药物反应分子靶标奠定基础。方法 日本血吸虫感染新西兰大白兔,吡喹酮治疗者为实验组,未经吡喹酮治疗者为对照组,应用PCR-Select cDNA Subtraction试剂盒进行SSH分析,分别构建正向和反向消减cDNA文库,对文库进行筛选,挑取阳性克隆测序获得差异表达EST或新EST,并进行生物信息学分析。结果 从2个消减文库中共筛选到39个有效阳性克隆,其中正向文库22个,反向文库17个,分析表明这些EST编码主要是一些酶及与蛋白质合成和降解相关的蛋白质。结论 成功构建日本血吸虫成虫吡喹酮治疗前后消减文库,为进一步研究吡喹酮治疗血吸虫病的分子机制奠定了基础。

关键词: 日本血吸虫成虫, 吡喹酮, 抑制性消减杂交, 差异表达序列

Abstract:

Objective To establish subtracted cDNA libraries using the suppression subtractive hybridization (SSH) technique based on the livers of New Zealand white rabbits infected with Schistosoma japonicum before and after praziquantel treatment, providing the basis for screening the drug-response molecular target during the therapy. Methods New Zealand white rabbits infected with S. japonicum were assigned to two groups to receive praziquantel medication or blank, respectively. The SSH technique was conducted to build the forward and reverse subtracted cDNA libraries using the PCR-Select cDNA Subtraction kit. The differentially expressed ESTs were screened and positive clones were isolated from the libraries for bioinformatic analysis.Results Thirty-nine positive clones were isolated from two subtracted cDNA libraries, including 22 from the forward library and 17 from the reverse library. The analysis revealed that the ESTs were mainly responsible for coding enzymes and proteins associated with protein synthesis and degradation. Conclusion The subtracted cDNA libraries before and after praziquantel treatment for S. japonicum were successfully constructed, laying a solid foundation for further research into the molecular mechanisms of this medication therapy.

Key words: Adult Schistosoma japonicum, Praziquantel, Suppression subtractive hybridization, Differentially expressed sequences

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