日本三角涡虫Dj-β-catenin-1基因干扰载体的构建与干扰效果的鉴定
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摘要
为了研究抑制Dj-β-catenin-1基因对涡虫再生的影响,构建了L4440-Dj-β-catenin-1干扰载体.将含有重组质粒L4440-Dj-β-catenin-1的HT115细菌经IPTG诱导后直接喂食涡虫.结果表明,Dj-β-catenin-1基因经RNA干扰后涡虫不能正常再生,面向后端伤口再生出一个头部而不是尾.此外,Dj-β-catenin-1基因的沉默还能使涡虫正常的尾转变成头.上述工作为进一步研究Dj-β-catenin-1基因在日本三角涡虫再生中的作用奠定良好基础.
In order to investigate the effects of inhibition of Dj-β-catenin-1 gene on planarian regeneration,the interference vector of L4440-Dj-β-catenin-1 is constructed in this research.After the IPTG induction of dsRNA,the bacteria HT115 containing the recombinant plasmid L4440-Dj β-catenin-1 are directly fed to planarians.The results show that RNA interference(RNAi)of Dj-β-catenin-1 causes the inappropriate regeneration of a head instead of a tail at posterior amputations.In addition,the silencing of Dj-β-catenin-1 is sufficient to transform the normal tail into a head.This work plays a good foundation for further studying the function of Dj-β-catenin-1 gene in planarian regeneration.
In order to investigate the effects of inhibition of Dj-β-catenin-1 gene on planarian regeneration,the interference vector of L4440-Dj-β-catenin-1 is constructed in this research.After the IPTG induction of dsRNA,the bacteria HT115 containing the recombinant plasmid L4440-Dj β-catenin-1 are directly fed to planarians.The results show that RNA interference(RNAi)of Dj-β-catenin-1 causes the inappropriate regeneration of a head instead of a tail at posterior amputations.In addition,the silencing of Dj-β-catenin-1 is sufficient to transform the normal tail into a head.This work plays a good foundation for further studying the function of Dj-β-catenin-1 gene in planarian regeneration.
引文
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[2]Su H,Suredagomez M,Rabanedalombarte N,et al.A C-terminally truncated form ofβ-catenin acts as a novel regulator of Wnt/β-catenin signaling in planarians[J].Plos Genetics,2017,13(10):e1007030.
[3]Nelson W J,Nusse R.Convergence of Wnt,bate-catenin,and cadherin pathways[J].Science,2004,303(5663):1483-1487.
[4]Clevers H,Nusse R.Wnt/β-catenin signaling and disease[J].Cell,2012,149(6):1192-1205.
[5]Ormanns S,Neumann J,Horst D,et al.WNT signaling and distant metastasis in colon cancer through transcriptional activity of nuclearβ-Catenin depend on active PI3Ksignaling[J].Oncotarget,2014,5(10):2999-3011.
[6]陈劲松,梁庆模.Wnt/β-Catenin信号通路与抗癌治疗新靶点[J].中国临床医学,2007,14(1):118-121.
[7]陈广文,马克学.淡水涡虫:分类地位、胚胎发生和再生[J].河南师范大学学报(自然科学版),2008,36(4):140-144.
[8]Reddien P W,Sánchez A A.Fundamentals of planarian regeneration[J].Annual Review of Cell&Developmental Biology,2004,20(1):725-757.
[9]Gurley K A,Rink J C,Alvarado A S.β-catenin defines head versus tail identity during planarian regeneration and homeostasis[J].Science,2008,319(5861):323-327.
[10]Petersen C P,Reddien P W.Smed-betacatenin-1is required for anteroposterior blastema polarity in planarian regeneration[J].Science,2008,319(5861):327-330.
[11]陆国清,王春玲,郝宇琼,等.转G10eve基因棉花的获得及草甘膦抗性的初探[J].棉花学报,2018,30(1):21-28.
[12]丁颜朋,葛晓阳,王鹏,等.棉花盐胁迫应答基因GhEXO70B1功能分析[J].棉花学报,2018,30(6):423-434.
[13]Iglesias M,Almuedocastillo M,Aboobaker A A,et al.Early planarian brain regeneration is independent of blastema polarity mediated by the Wnt/β-catenin pathway[J].Developmental Biology,2011,358(1):68-78.
[14]Hill E M,Petersen C P.Wnt/Notum spatial feedback inhibition controls neoblast differentiation to regulate reversible growth of the planarian brain[J].Development,2015,142(24):4217-4229.
[15]Kim J K,Gabel H W,Kamath R S,et al.Functional genomic analysis of RNA interference in C.elegans[J].Science,2005,308(5725):1164-1167.