›› 2011, Vol. 23 ›› Issue (4): 0-711.

• 动物科学 •    

应用荧光定量PCR对日本血吸虫miRNAs表达量的研究

郝力力1 ,李 锐2,郑 威3,*   

  1. 1西南民族大学 生命科学与技术学院, 四川 成都 610041;2浙江省农业科学院 农产品质量标准研究所, 浙江 杭州 310021;3中国农业科学院 广西水牛研究所, 广西 南宁 530001
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-07-25 发布日期:2011-07-25

Quantification analysis of miRNAs expression level in different developing stages of Schistosoma japonicum using real time PCR

HAO Li-li;LI Rui ;ZHENG Wei;*   

  1. 1College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China;2Institute of Quality and Standard for Agricultural Products, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021,China;3Guangxi Buffalo Research Institute,Chinese Academy of Agricultural Science, Nanning 530001,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-07-25 Published:2011-07-25

摘要: 近来通过Solexa高通量测序鉴定出了大量日本血吸虫的miRNAs序列,为确定这些miRNAs在日本血吸虫不同生活史阶段的表达量,采用SYBR染料法和Race试剂盒相结合的荧光定量方法对虫卵、尾蚴、14 d童虫、雄虫、雌虫阶段的miRNAs表达量进行了检测分析,发现虫体不同阶段miRNAs表达差异巨大,其中虫卵阶段表达量最低,而虫体其他阶段有着各自的表达特点,同时也用Northern对荧光定量结果进行了进一步的验证。结果表明,这些miRNAs可能在日本血吸虫不同发育阶段的基因转录调控中起着重要作用。

关键词: 日本血吸虫, miRNAs, 荧光定量PCR

Abstract: At present, many new miRNAs in Schistosoma japonicum are discovered by high\|throughout sequencing technology such as Solexa. To investigate the expression level of these miRNAs in different life stages of Schistosoma japonicum, real-time PCR was used to relatively quantify the dominantly expressed miRNAs in egg, cercarium, schistosomulum, male and female adult by combination of SYBR Green PCR and Race kit. It was found that these miRNAs were highly expressed at stages other than the egg and with different expression pattern in other stages, which were further confirmed by Northern blot. In summary, the results suggested that miNRAs might play an important role in regulation of genes transcription in different developing stages of Schistosoma japonicum.

Key words: Schistosoma japonicum, miRNAs, real time PCR