Identification and RNAi of a Bursaphelenchus xylophilus dauer formation gene: Bx-DAF6
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摘要: 为探究松材线虫抗逆态幼虫形成的分子机理,本文对抗逆态基因daf-6进行了鉴定和基因沉默研究。应用BLAST比对技术,在松材线虫基因组数据中鉴定得到秀丽线虫daf-6的同源基因,命名为Bx-DAF6。应用PCR技术进行Bx-DAF6完整蛋白编码区(CDS)扩增。采用RNAi技术进行Bx-DAF6沉默,分析该基因的沉默对松材线虫抗逆态幼虫形成的影响。PCR扩增得到CDS区全长2 700 bp,Bx-DAF6编码的蛋白质具有固醇传感结构域。Bx-DAF6沉默抑制了2龄幼虫转变为抗逆态幼虫,表明Bx-DAF6在2龄幼虫转变为抗逆态幼虫生理过程中起正向调控作用。Abstract: To explore the molecular mechanism of the Bursaphelenchus xylophilus specialized third-stage dauer larva formation, we studied the Bx-DAF6 gene using the RNAi method. Based on the B. xylophilus genome, Bx-DAF6 was identified by the method of BLAST, and cloned by PCR. The gene function was studied by the suppression of Bx-DAF6 transcript in combination with the RNAi method. The results indicated that the Bx-DAF6 CDS was 2 700 bp, which encodes a protein with a domain homologous to sterol-sensing. The suppression of the Bx-DAF6 transcript by RNAi led to a reduction in the number of specialized third-stage dauer larva formation. Our results indicated that Bx-DAF6 regulated positively the formation of specialized third-stage dauer larvae.
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