College of Animal Science & Technology, Shihezi University, Shihezi, Xinjiang 832000, China.
Chin Med J (Engl). 2011 Sep;124(18):2838-44.
Cystic echinococcosis due to Echinococcus granulosus (E. granulosus) is one of the most important chronic helminthic diseases, especially in sheep/cattle-raising regions. The larval stage of the parasite forms a cyst that grows in the liver, lung, or other organs of the host. To ensure a long life in the host tissues, the parasite establishes complex inter-cellular communication systems between its host to allow its differentiation toward each larval stage. Recent studies have reported that this communication is associated with the extracellular signal-regulated kinase (ERK) mitogen-activated protein kinase cascade in helminth parasites, and in particular that these protein kinases might serve as effective targets for a novel chemotherapy for cystic echinococcosis. The aim of the present study investigated the biological function of a novel ERK ortholog from E. granulosus, EgERK.
DNA encoding EgERK was isolated from protoscolices of E. granulosus and analyzed using the LA Taq polymerase chain reaction (PCR) approach and bioinformatics. Reverse transcription PCR (RT-PCR) was used to determine the transcription level of the gene at two different larval tissues. Western blotting was used to detect levels of EgERK protein. The expression profile of EgERK in protoscolices was examined by immunofluorescence.
We cloned the entire Egerk genomic locus from E. granulosus. In addition, two alternatively spliced transcripts of Egerk, Egerk-A, and Egerk-B were identified. Egerk-A was found to constitutively expressed at the transcriptional and protein levels in two different larval tissues (cyst membranes and protoscolices). Egerk-A was expressed in the tegumental structures, hooklets, and suckers and in the tissue surrounding the rostellum of E. granulosus protoscolices.
We have cloned the genomic DNA of a novel ERK ortholog from E. granulosus, EgERK (GenBank ID HQ585923), and found that it is constitutively expressed in cyst membrane and protoscolex. These findings will be useful in further study of the biological functions of the gene in the growth and development of Echinococcus and will contribute to research on novel anti-echinococcosis drug targets.
由细粒棘球绦虫（Echinococcus granulosus，E. granulosus）引起的包虫病是最重要的慢性寄生虫病之一，尤其在绵羊/牛养殖区。寄生虫的幼虫阶段形成囊肿，在宿主的肝脏、肺部或其他器官中生长。为了在宿主组织中保证长期生存，寄生虫在其与宿主之间建立了复杂的细胞间通讯系统，以允许其向每个幼虫阶段分化。最近的研究表明，这种通讯与寄生虫细胞外信号调节激酶（ERK）丝裂原激活蛋白激酶级联反应有关，特别是这些蛋白激酶可能成为包虫病新化疗的有效靶点。本研究旨在研究来自细粒棘球绦虫的新型 ERK 同源物 EgERK 的生物学功能。
使用 LA Taq 聚合酶链反应（PCR）方法和生物信息学从细粒棘球蚴原头节中分离编码 EgERK 的 DNA，并进行分析。逆转录 PCR（RT-PCR）用于确定该基因在两种不同幼虫组织中的转录水平。Western 印迹用于检测 EgERK 蛋白的水平。通过免疫荧光检测 EgERK 在原头节中的表达谱。
我们从细粒棘球绦虫中克隆了整个 Egerk 基因组基因座。此外，还鉴定了 Egerk 的两种选择性剪接转录本，Egerk-A 和 Egerk-B。Egerk-A 在两种不同的幼虫组织（囊膜和原头节）中在转录和蛋白水平上均持续表达。Egerk-A 在细粒棘球蚴原头节的表皮结构、钩状结构、吸盘和口盖周围的组织中表达。
我们从细粒棘球绦虫中克隆了一种新型 ERK 同源物 EgERK（GenBank ID HQ585923）的基因组 DNA，并发现它在囊膜和原头节中持续表达。这些发现将有助于进一步研究该基因在细粒棘球蚴生长发育中的生物学功能，并为新型抗包虫病药物靶点的研究做出贡献。
