论文部分内容阅读
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PCRSelectcDNASubtraction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。
OBJECTIVE: To establish a subtractive subtracted cDNA library of praziquantel against Schistosoma japonicum infection in New Zealand white rabbits by suppression subtractive hybridization (SSH) technique, and to lay a foundation for screening molecular targets of in vivo drug reactions in Schistosoma japonicum during the course of praziquantel treatment. Methods New Zealand white rabbits were infected with Schistosoma japonicum and praziquantel was used as the experimental group. The control group was treated with praziquantel. SSH analysis was carried out by using PCRSelect cDNAsubtraction kit to construct forward and reverse subtracted cDNA library, The positive clones were selected and sequenced to obtain differentially expressed ESTs or novel ESTs and analyzed by bioinformatics. Results A total of 39 valid positive clones were screened from two subtractive libraries, of which 22 were forward and 17 were reverse transcripts. Analysis showed that these ESTs were mainly encoded by enzymes and proteins involved in protein synthesis and degradation. Conclusion The successful construction of Schistosoma japonicum adult praziquantel treatment before and after subtraction library, in order to further study the molecular mechanism of praziquantel treatment of schistosomiasis laid the foundation.