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Expression and functional characterization of a Rho-family small GTPase CDC42 from Trichinella spiralis

机译:旋毛虫的Rho家族小GTPase CDC42的表达和功能表征

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摘要

A full-length cDNA encoding a Rho-family small GTPase gene cdc42 of Trichinella spiralis was expressed in E. coli. The recombinant protein of TsCDC42 was purified and used to raise the polyclonal antibodies. The expression of TsCDC42 in different stages of parasite was investigated. The western blot showed that TsCDC42 was expressed in all stages of T. spiralis, including newborn larvae, muscle larvae and adult worms. The immuno-localization revealed that TsCDC42 was ubiquitously distributed in the newborn larvae, muscle larvae and adult worm. Cross-species RNAi was done by knockdown Tscdc42 RNAi in C. elegans. The results revealed that endogenous expression level of CDC42 was decreased by knockdown Tscdc42 RNAi in C. elegans, and this knockdown reduced the progeny of C. elegans. It suggested that Tscdc42 might play the same roles in the early development of T. spiralis.
机译:在大肠杆菌中表达编码旋毛虫的Rho家族小GTPase基因cdc42的全长cDNA。纯化TsCDC42的重组蛋白,并用于产生多克隆抗体。研究了TsCDC42在寄生虫不同阶段的表达。 Western印迹表明,TsCDC42在螺旋藻的所有阶段均有表达,包括新生幼虫,肌肉幼虫和成虫。免疫定位表明,TsCDC42普遍分布在新生幼虫,肌肉幼虫和成虫中。通过敲除秀丽隐杆线虫中的Tscdc42 RNAi,完成了跨物种RNAi。结果显示,敲除线虫中的Tscdc42 RNAi可降低CDC42的内源表达水平,而这种敲除可减少线虫的后代。这表明Tscdc42可能在螺旋藻的早期发育中发挥相同的作用。

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