IDENTIFYING ANTIGENIC PROTEINS OF BARTONELLA HENSELAE

鉴定汉塞巴尔通体的抗原蛋白

基本信息

批准号：
6378431
负责人：
MICHAEL W SCHMIEDERER
金额：
$ 2.12万
依托单位：
UNIVERSITY OF SOUTH FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-06-01 至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/6378431
关键词：
Escherichia coli Rickettsiales SDS polyacrylamide gel electrophoresis antigens bacteria infection mechanism bacterial antigens bacterial genetics diagnosis design /evaluation enzyme linked immunosorbent assay gene mutation genetic library human tissue immunoprecipitation nucleic acid sequence opportunistic infections polymerase chain reaction protein purification protein structure recombinant proteins serology /serodiagnosis western blottings

Escherichia coli Rickettsiales SDS polyacrylamide gel electrophoresis antigens bacteria infection mechanism bacterial antigens bacterial genetics diagnosis design /evaluation enzyme linked immunosorbent assay gene mutation genetic library human tissue immunoprecipitation nucleic acid sequence opportunistic infections polymerase chain reaction protein purification protein structure recombinant proteins serology /serodiagnosis western blottings

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项目摘要

DESCRIPTION: Applicant's Abstract Bartonella henselae is an opportunistic pathogen that causes disease in AIDS patients and intravenous drug abusers. Diagnosis of disease would be enhanced if a specific antigen could be identified. To this end the following aims have been identified. Aim 1: To identify genes encoding major antigens of B. henselae. A DNA lambda phage library will be screened with HIV-1 infected patient sera that are positive by indirect fluorescence assay (IFA) to Bartonella spp. Clones will be rescued and expressed in E. Coli. Aim 2: Determine the nucleotide sequence and control mechanisms of the gene coding for the antigen. Vector specific primers and exonuclease digestion of the insert will be used with dideoxynucleotide chain termination sequencing. Aim 3: Evaluate the diagnostic potential of the recombinant protein and its associated gene. Purified protein will be used as an antigen in ELISA assay. Aim 4: Determine the location, arrangement, and function of the antigen. Mutations of the gene will be transferred to B. henselae by conjugation with E. Coli.

描述：申请人的摘要 Bartonella henselae是一种机会性病原体，会导致艾滋病疾病患者和静脉吸毒者。诊断疾病将是如果可以识别特定的抗原，可以增强。为此结束已经确定了以下目标。目标1：识别编码的基因 Henselae的主要抗原。将筛选DNA lambda噬菌体库与HIV-1感染的患者血清，间接荧光为阳性分析（IFA）到Bartonella spp。克隆将在E中获救和表达。大肠杆菌。目标2：确定核苷酸序列和控制机制抗原编码的基因。矢量特定引物和外切酶插入物的消化将与二乙氧基核苷酸链一起使用终止测序。目标3：评估重组蛋白及其相关基因。将使用纯化的蛋白质作为ELISA测定法中的抗原。目标4：确定位置，安排，和抗原的功能。基因的突变将转移到 B. henselae与大肠杆菌结合。