A study on exocrine substances of Trichinella that induce dedifferentiation and transformation of infected muscle cell

旋毛虫外分泌物质诱导感染肌细胞去分化和转化的研究

• 批准号: 09670255
• 负责人: TAKAHASHI Yuzo
• 金额: $ 1.86万
• 依托单位: Gifu University School of Medicine
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09670255/
• 关键词: Trichinella; ES products; stichosome; peptide; 筋肉細胞; 分化

Mass and pure production of peptide using cDNA library of Trichinella was achieved for the two reasons. One is to obtain antigen for immunodiagnostic purposes and/or immunoresponse analysis. The other is to obtain and analysis protein that may alter gene expression of host muscle cells, known as nurse cell formation. We have constructed cDNA library of T spiralis and T pseudospiralis, and produced some of possibly important peptides. The method includes isolation of mRNA from muscle larvae, cDNA library construction, immunoscreening of E coli colony against infected sera. Ten-positive clones were obtained, 5 out of which were sequenced. Homology analysis showed Clone 111 has homology to protenase inhibitor, and Clone 132 seemed to be identical to one reported by Despommier. Other clones were unique showing no similarity to previously reported sequences. Western blot analysis of synthesized peptides reacted against anti ES antibody or infected sera, or failed to react.

利用旋毛虫cDNA文库实现肽的大规模和纯生产有两个原因。一是获得用于免疫诊断目的和/或免疫反应分析的抗原。另一个是获取和分析可能改变宿主肌肉细胞基因表达的蛋白质，称为护士细胞形成。我们构建了螺旋毛虫和假螺旋毛虫的cDNA文库，并产生了一些可能重要的肽。该方法包括从肌肉幼虫中分离mRNA、构建cDNA文库、针对感染血清对大肠杆菌菌落进行免疫筛选。获得了 10 个阳性克隆，其中 5 个进行了测序。同源性分析显示克隆111与蛋白酶抑制剂具有同源性，克隆132似乎与Despommier报道的克隆相同。其他克隆是独特的，与之前报道的序列没有相似性。合成肽的蛋白质印迹分析与抗ES抗体或感染的血清发生反应，或未发生反应。

项目成果

Xu, Takahashi: "Electron microscopic observations of the stichosome during the normal development of Trichinella spiralis" Journal of Electron Microscopy. 46. 439-442 (1997)

Xu，Takahashi：“旋毛虫正常发育过程中针体的电子显微镜观察”电子显微镜杂志。

Z.WU, I.NAGANO' Y.TAKAHASHI: "Differences and similarities between Teichinella spiralis and T,pseudospiralis" Parasitology. 116. 61-66 (1998)

Z.WU、I.NAGANO Y.TAKAHASHI：“Teichinellaspiralis 和 T,pseudospiralis 之间的差异和相似之处”寄生虫学。

Hisao Yoshikawa: "Genomic polymorphism among Blastocystis hominis strains and development of subtype-specific diagnostic primers." Molecular and Cellular Probes. 12. 153-159 (1998)

Hisao Yoshikawa：“人芽囊藻菌株的基因组多态性和亚型特异性诊断引物的开发。”

Nagano et al: "Detection of verotoxin-Producing E.Col :" Micobiol.Immurnol. 42. 371-376 (1998)

Nagano 等人：“检测产生维罗毒素的大肠杆菌：”Micobiol.Immurnol。

Z.WU: "The detection of Trichinella with polymerase chain reaction (PCR) primers constructed using sequences of random amplified polymorphic DNA (RAPD) or sequences of complementary DNA encoding excretory-secretory- (E-S) glycoproteins." Parasitology. 117

Z.WU：“使用随机扩增多态性 DNA (RAPD) 序列或编码排泄分泌 (E-S) 糖蛋白的互补 DNA 序列构建的聚合酶链反应 (PCR) 引物检测旋毛虫。”