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在人胚胎干细胞中高效表达PELO蛋白
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作者 胡旭林 纪家葵 《生物技术通报》 CAS CSCD 北大核心 2013年第10期170-176,共7页
以PELO全长cDNA为模板,采用TOPO克隆的方法,将人的PELO基因克隆到pENTR-D-TOPO载体上,形成pENTR-D-PELO克隆载体。然后通过同源重组将含有EF1α启动子的pENTR-5’-EF1α和pENTR-D-PELO重组到表达载体p2k7上,形成EF1α-PELO-p2k7表达载... 以PELO全长cDNA为模板,采用TOPO克隆的方法,将人的PELO基因克隆到pENTR-D-TOPO载体上,形成pENTR-D-PELO克隆载体。然后通过同源重组将含有EF1α启动子的pENTR-5’-EF1α和pENTR-D-PELO重组到表达载体p2k7上,形成EF1α-PELO-p2k7表达载体。将构建好的PELO高表达载体在293FT细胞中制备成相应的慢病毒表达载体,并用慢病毒侵染人胚胎干细胞,最终成功实现在人胚胎干细胞中稳定且高效地表达PELO蛋白。 展开更多
关键词 pelo 高表达 胚胎干细胞 慢病毒 同源重组
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The mRNA surveillance factor Pelo restricts rice virus propagation in insect vectors and host plants
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作者 Xinyan Sun Heran Guan +1 位作者 Taiyun Wei Dongsheng Jia 《Phytopathology Research》 2024年第1期482-493,共12页
Many devastating plant viruses are transmitted by insect vectors among plant hosts in a persistent-propagative manner.Pelota(Pelo)is an evolutionarily conserved protein involved in the mRNA surveillance system.In this... Many devastating plant viruses are transmitted by insect vectors among plant hosts in a persistent-propagative manner.Pelota(Pelo)is an evolutionarily conserved protein involved in the mRNA surveillance system.In this study,it was found that the accumulation of Pelo proteins are slightly decreased during the propagation of the fijivirus southern rice black-streaked dwarf virus(SRBSDV)in rice and transmission vector planthopper(Sogatella furcifera).The tubular protein P7-1 encoded by SRBSDV interacted with Pelo of rice or planthopper vector.Overexpression or knockdown of Pelo expression inhibits the formation of P7-1 tubules in insect cells,further exerting antiviral activity.Furthermore,overexpression or knockout of Pelo expression in transgenic rice plants also inhibits the effective propagation of SRBSDV as well as two other rice viruses of different families.The slight reduction of Pelo accumulation during SRBSDV propagation in rice and insect vectors would avoid Pelo-mediated excessive inhibition of P7-1 tubule formation,ensuring effective virus propagation.Our findings provide insights into how the up-or down-regulated expression of Pelo in rice hosts and insect vectors elevate their resistance to rice viruses. 展开更多
关键词 mRNA surveillance system pelo Rice virus RESISTANCE
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