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干扰日本血吸虫凋亡抑制因子最佳siRNA分子的筛选
  • ISSN号:1002-2694
  • 期刊名称:《中国人兽共患病学报》
  • 时间:0
  • 分类:R383.2[医药卫生—医学寄生虫学;医药卫生—基础医学]
  • 作者机构:[1]中国农业科学院上海兽医研究所,农业部动物寄生虫学重点开放实验室,上海200241
  • 相关基金:国家自然科学基金(No.30901068)资助
中文摘要:

目的筛选干扰日本血吸虫凋亡抑制因子(Inhibitor of apoptosis protein of Schistosoma japonicum,SjIAP)较好的siRNA分子。方法应用生物信息学设计了针对sjIAP的3对siRNA分子。利用脂质体将每对siRNA分子和表达IAP的重组质粒传染到HEK293T细胞中,利用实时定量RT—PCR和免疫印迹检测IAP的表达。将siRNA分子与日本血吸虫童虫(12d)进行体外共培养,利用实时定量RT—PCR和免疫印迹检测虫体内的lAP表达。结果细胞转染实验的实时定量RT—PCR和免疫印迹分析表明本研究获得了2个干扰sjIAP较好的siRNA分子,且其中1对siRNA分子可显著抑制日本血吸虫虫体内IAP蛋白的表达,同时虫体Caspase活性有所上升。结论本研究获得了2个干扰日本血吸虫lAP较好的siRNA分子,为进一步利用RNA干扰研究血吸虫lAP的功能奠定了前期基础。

英文摘要:

Our previous study suggests that the inhibitor apoptosis protein in Schistosoma japonicum (SjlAP) may play important roles in parasitic living and development as well as in the host parasite interactions. To obtain the best efficacy of small interfering RNA (siRNA) for silencing the SjIAP, three siRNA duplexes targeting SjlAP were bioinformatically de signed and chemical synthesized. Then, each siRNA duplex and the recombinant plasmid for expressing SjIAP were co-trans fected into human HEK293T cells. Real-time RT-PCR and Western blot analyses indicated that the siRNA 951 showed the best efficacy for silencing SjlAP expression in both mRNA and protein levels. Then, the siRNA 95l was further added into the me dium of in vitro cultured schistosomula (12 days). Western blot analysis indicated that the expression of SjlAP to some extent inhibited at the protein level. In addition, the activities of Caspase 3/7 were also increased in the worms treated with siRNA 951. In conclusion, the best efficacy of siRNA for silencing SjlAP was obtained and the present study provides foundational in- formation for the further investigation on SjIAP functions by using RNA interference.

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期刊信息
  • 《中国人兽共患病学报》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学技术协会
  • 主办单位:中国微生物学会
  • 主编:严延生
  • 地址:福州市津泰路76号
  • 邮编:350001
  • 邮箱:rsghb@fjcdc.com.cn
  • 电话:0591-87552018
  • 国际标准刊号:ISSN:1002-2694
  • 国内统一刊号:ISSN:35-1284/R
  • 邮发代号:34-46
  • 获奖情况:
  • 中国基础医学核心期刊,中国生物医学核心期刊,中国科学引文数据库列为来源期刊及统计源,中国期刊方阵“双效”期刊
  • 国内外数据库收录:
  • 俄罗斯文摘杂志,美国化学文摘(网络版),英国农业与生物科学研究中心文摘,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:9000