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首页> 外文期刊>The Journal of Experimental Biology >REPORTER GENE CONSTRUCTS SUGGEST THAT THE CAENORHABDITIS ELEGANS AVERMECTIN RECEPTOR BETA-SUBUNIT IS EXPRESSED SOLELY IN THE PHARYNX
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REPORTER GENE CONSTRUCTS SUGGEST THAT THE CAENORHABDITIS ELEGANS AVERMECTIN RECEPTOR BETA-SUBUNIT IS EXPRESSED SOLELY IN THE PHARYNX

机译：报道基因表明，食管念珠菌类阿维菌素受体β-亚单位仅在咽部表达

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Gene promoter/LacZ reporter constructs were made in order to analyse the expression of the beta-subunit of the Caenorhabditis elegans glutamate-gated Cl- channel (Glu-Cl) receptor. Southern blot analysis of the C. elegans cosmid C35E8 identified a 4kbp EcoRI fragment which contained the 5' portion of the Glu-Cl beta coding sequence together with 5' flanking sequences. This was subcloned and used as the template for polymerase chain reaction (PCR) amplification of a DIVA fragment encoding the first 24 amino acid residues of Glu-Cl beta together with 1.4 kbp of 5' genomic sequence. The fragment was subcloned into the LacZ expression vector pPD22.11 to form a translational reporter fusion. After injection of the construct into worms, six stably transformed lines were established and assayed for beta-galactosidase activity. Stained nuclei were observed in the pharyngeal metacorpus in adults and in all larval stages, and stained nuclei were seen in many embryos undergoing morphogenesis. Additional stained nuclei towards the terminal bulb of the pharynx were observed in larval stages. These results provide further evidence that the Glu-Cl receptor mediates the glutamatergic inhibition of pharyngeal muscle via the M3 motor neurone and point to inhibition of pharyngeal pumping as a major mode of action for avermectins. [References: 40]
机译：制备基因启动子/ LacZ报道基因构建体是为了分析秀丽隐杆线虫谷氨酸门控的Cl-通道（Glu-Cl）受体的β-亚基的表达。秀丽隐杆线虫粘粒C35E8的Southern印迹分析鉴定出4kbp EcoRI片段，其包含Glu-C1β编码序列的5'部分以及5'侧翼序列。将其亚克隆并用作模板，用于DIVA片段的聚合酶链反应（PCR）扩增，该片段编码Glu-C1 beta的前24个氨基酸残基以及1.4 kbp的5'基因组序列。将该片段亚克隆到LacZ表达载体pPD22.11中，形成翻译报告基因融合体。将构建体注射到蠕虫中后，建立了六个稳定转化的品系，并测定了β-半乳糖苷酶的活性。在成虫和所有幼虫的咽部咽部中均观察到染色的细胞核，并且在经历形态发生的许多胚胎中均观察到染色的细胞核。在幼体阶段观察到朝向咽部终末球的其他染色核。这些结果提供了进一步的证据，表明Glu-Cl受体通过M3运动神经元介导了咽肌对谷氨酸的抑制作用，并表明抑制咽抽动是阿维菌素的主要作用方式。 [参考：40]

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