CLONING AND EXPRESSION OF THE HOOKWORM ANTICOAGULANT

钩虫抗凝剂的克隆和表达

• 批准号: 2886004
• 负责人: MICHAEL CAPPELLO
• 金额: $ 9.53万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-15 至 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2886004
• 关键词: Baculoviridae; Escherichia coli; Nematoda; activation product; anticoagulants; blood coagulation; coagulation factor IX; coagulation factor VII; coagulation factor X; complementary DNA; genetic library; immunocytochemistry; in situ hybridization; monoclonal antibody; nucleic acid sequence; polymerase chain reaction; protease inhibitor; protein purification; protein sequence; protein structure function; recombinant proteins; thromboplastin; tissue /cell culture; transfection /expression vector; western blottings

DESCRIPTION: Phase I of the planned program consists of taking a summer course in molecular biology and auditing five courses. Phases I and II will include different portions of the research program. Human hookworm infection is characterized by chronic GI bleeding and iron deficiency anemia. The adult worm facilitates blood feeding through secretion of a potent anticoagulant called the Hookworm Extrinsic Pathway Inhibitor (HEPI). The candidate has already isolated this novel 16 kDa protein from lysates of Ancylostoma duodenale. Synthetic oligonucleotides from amino acid sequences generated from tryptic digests of the protein have been used to generate and clone a PCR product containing a partial coding region sequence. The candidate plans to clone the full length gene from a cDNA library, to sequence this and to express recombinant HEPI in a bacterial or mammalian vector system. This expressed protein will be used to generate antibody for immunolocallization of the protein in the organism and for biochemical assays of protein activity.

描述：计划项目的第一阶段包括一个夏天

项目成果

Ancylostoma caninum anticoagulant peptide-5: immunolocalization and in vitro neutralization of a major hookworm anti-thrombotic.

• DOI: 10.1016/s0166-6851(01)00276-6
• 发表时间: 2001-06
• 期刊: Molecular and biochemical parasitology
• 影响因子: 1.5
• 作者: L. Harrison;J. L. Córdova;M. Cappello