一种基于干扰肌动蛋白基因导致方形网纹溞(Ceriodaphnia quadrangula)死亡的RNA干扰(RNAi)模型构建  

A RNAI MODEL CONSTRUCTION BASED ON ACTIN GENE INTERFERENCE INDUCING DEATH OF CERIODAPHNIA QUADRANGULA

作　　者：陈宏健[1] 谢松[1,2] 李理想 柳峰松[1,2] 

机构地区：[1]河北大学生命科学学院,保定071002 [2]河北省无脊椎动物系统学与应用实验室,保定071002

出　　处：《海洋与湖沼》2014年第2期322-327,共6页Oceanologia Et Limnologia Sinica

基　　金：河北省自然科学基金项目;C2010000256号;C2011201027号;教育部高等学校博士学科点专项科研基金项目;20101301120005

摘　　要：方形网纹溞(Ceriodaphnia quadrangula)可作为研究水生动物生物学的良好模式动物。为探索细菌介导表达的双链RNA(dsRNA)对方形网纹溞进行RNA干扰(RNAi)技术是否可行,本研究构建了方形网纹溞肌动蛋白(actin)基因的干扰载体Actin-L4440,并将其转入大肠杆菌HT115。此宿主菌经异丙基-β-D-硫代吡喃半乳糖苷(IPTG)诱导后能表达actin基因部分片段的dsRNA。将此宿主菌投喂方形网纹溞,能抑制其体内actin基因的表达,并导致方形网纹溞死亡率明显提高。本研究成功构建了方形网纹溞的RNAi模型,为以后的研究提供了一定的参考。We developed a method of RNA interference based on bacterial-mediated expressed dsRNA in Ceriodaphnia quadrangular. A part of actin gene fragment was inserted into L4440 dsRNA expression vector. Then the recombinant plasmid was constructed and transformed to Escherichia coli HT115. The dsRNA was expressed after IPTG induction. Quantitative PCR data showed that the actin gene expression was knocked down after feeding and soaking in E. coli HT115 with actin-L4440 recombinant plasmid. Meanwhile, knocking down the actin expression led to an increase in the mortality of Ceriodaphnia quadrangula. Thirty-six hours after feeding, the mortality of RNAi groups(71.67%) nearly doubled that of the control(37.78%). Almost all C. quadrangular in RNAi group were dead 48 hours after feeding. Therefore, the model of RNAi for C. quadrangular is successfully constructed that may offer a reference for in-depth research in the future.

关 键 词：RNA干扰 方形网纹溞(Ceriodaphnia quadrangula) 肌动蛋白 

分 类 号：Q78[生物学—分子生物学]