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首页> 外文期刊>Veterinary Parasitology >Isolation and characterization of a gene encoding carbonic anhydrase from Ostertagia ostertagi and quantitative measurement of expression during in vivo exsheathment.
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Isolation and characterization of a gene encoding carbonic anhydrase from Ostertagia ostertagi and quantitative measurement of expression during in vivo exsheathment.

机译:分离和鉴定Ostertagia ostertagi碳酸酐酶编码基因,并在体内放出过程中定量测量表达。

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The first event in the establishment of Ostertagia ostertagi infection in cattle is exsheathment. Exsheathment is the process whereby the L(2) cuticle retained from the previous molt is cast from the L3. For those trichostrongyle nematode species with a predilection site in the abomasum, such as O. ostertagi, exsheathment is initiated as the larvae pass through the rumen. Although the stimulus for exsheathment is not known, previously reported biochemical studies suggest a major role for the enzyme carbonic anhydrase (CA). Partial support for this hypothesis comes from the reported failure of the Haemonchus contortus L3 to exsheath following pretreatment with ethoxzolamide, a known inhibitor of CAs. Although convincing, a CA has not been previously reported from a trichostrongylid nematode. Therefore, our objective was to isolate a CA gene from O. ostertagi L3 and begin initial characterization studies. This work resulted in the successful isolation, cloning and sequencing of the first CA isolated from a gastrointestinal nematode. The gene, designated OoCA, shows 90.5% sequence identity with the CA eukaryotic consensus sequence, 78% similarity to the Caenorhabiditis elegans cah-6 and 55% similarity to the human CAIII. Sequence analysis of the genomic DNA encoding OoCA shows 8 exons and 7 introns covering 4.5kb. The first 1758 bases of the promoter region suggest OoCA may be regulated in part by transcription factors associated with hypoxic signaling and development. The mRNA profile of OoCA in exsheathing O. ostertagi L3 suggests this particular CA may play a role in immediate early developmental events following exsheathment initiation.
机译:在牛中建立Ostertagia ostertagi感染的第一个事件是放血。剥皮是从L3浇铸前一蜕皮残留的L(2)角质层的过程。对于在嗜好菌中有好发位点的那些线虫的线虫的线虫物种,例如O. ostertagi,当幼虫通过瘤胃时就开始着鞘。尽管不知道用于刺激的刺激作用,但是先前报道的生化研究表明碳酸酐酶(CA)的主要作用。对这一假说的部分支持来自于报道的乙型肝炎链球菌(Haemonchus contortus L3)在用已知的CAs抑制剂乙氧唑酰胺预处理后无法脱皮。尽管有说服力,但先前尚未从毛线虫线虫的线虫中报道过CA。因此,我们的目的是从O. ostertagi L3中分离出一个CA基因并开始初步的表征研究。这项工作成功分离,克隆和测序了从胃肠道线虫中分离出的首个CA。该基因命名为OoCA,与CA真核共有序列具有90.5%的序列同一性,与秀丽隐杆线虫cah-6有78%的相似性,与人CAIII有55%的相似性。编码OoCA的基因组DNA的序列分析显示8个外显子和7个内含子,覆盖4.5kb。启动子区域的前1758个碱基表明OoCA可能部分受与缺氧信号传导和发育相关的转录因子调控。 OoCA在外皮O. ostertagi L3中的mRNA谱表明,这种特定的CA可能在外皮启动后立即的早期发育事件中起作用。

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