DNA ANALYSIS OF PARASITES

寄生虫的 DNA 分析

• 批准号: 3960564
• 负责人: T E NASH
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF ALLERGY AND INFECTIOUS DISEASES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3960564
• 关键词: DNA; Giardia; endonuclease; epidemiology; genetic manipulation; genetic translation; giardiasis; microorganism genetics; molecular cloning; nucleic acid sequence; parasitic disease diagnosis; protozoal antigen

Nine new isolates of Giardia have been axenized and are in the process of being studied. The DNA of other isolates were analyzed by endonuclease restriction analysis and found to be different from most of the previously defined Giardia isolates. In order to study the structure and gene organization of the major 170kD antigen of isolate WB, the gene was cloned into an expression vector and a number of clones isolated. Sequencing of this gene is in progress. The insert from this vector was cloned into M13 and used as a probe in restriction endonuclease studies of various isolates. The gene is present in isolates that do not express the 170kD antigen. Mutants of Giardia could not be produced by commonly used procedures. Transfection of Giardia was attempted with Icarus hs neo gene which confers resistance to G418. Although RNA transcripts were found, no detectable translated products were found. The heat sock gene (hs-p) of Giardia was isolated and cloned into Puc18 and the promotor for the gene will be used with the neo gene. Translation products of RNAs from different isolates showed subtle but definite differences. A 43kD product was recognized by infected human and gerbil sera.

九个新的贾第虫分离株已被axenized，并正在进行 被研究。 用核酸内切酶对其他分离株的DNA进行分析 限制性内切酶分析，发现与以前的大多数不同， 定义的贾第虫分离株。 为了研究其结构和基因， 根据WB株主要170 kD抗原的结构，克隆了该基因 转化到表达载体中并分离出许多克隆。 测序 这个基因正在进行中。 将来自该载体的插入片段克隆到M13中 并用作各种限制性内切酶研究中的探针 分离株 该基因存在于不表达170 kD 抗原的 通常使用的方法不能产生贾第虫的突变体。 尝试用Icarus hs neo基因转染贾第虫， 抗G418 虽然发现了RNA转录物，但没有检测到 发现了翻译产物。贾第虫的热袜基因（hs-p）是 分离并克隆到Puc 18中，并使用该基因的启动子 neo基因。 来自不同分离株的RNA翻译产物显示出微妙的，但 明确的差异。 感染者可识别一个43 kD的产物， 沙鼠血清。