Nilsen T W, Maroney P A, Goodwin R G, Perrine K G, Denker J A, Nanduri J, Kazura J W
Department of Molecular Biology and Microbiology, Case Western Reserve University, School of Medicine and University Hospitals, Cleveland, OH 44106.
Proc Natl Acad Sci U S A. 1988 May;85(10):3604-7. doi: 10.1073/pnas.85.10.3604.
To facilitate biochemical studies of protective filarial antigens, a lambda gt11 cDNA library was constructed from Brugia malayi adult mRNA and screened with rabbit sera that recognizes a limited set of filarial antigens of approximately 25, 42, 60, and 112 kDa. Antigens of approximately equal to 25 and approximately equal to 60 kDa have been shown previously to induce enhanced clearance of microfilaremia in mice. A 154-base pair clone detected by immunological reactivity was used to isolate by hybridization a nearly full-length cDNA clone of 1.8 kilobases. Nucleotide-sequence analysis indicated that this clone was derived from a mRNA encoding a 63-kDa antigen. A fusion polypeptide containing 37 kDa of the Escherichia coli TrpE protein (anthranilate synthase) and 55 kDa of the cloned protein was recognized in immunoblot experiments with antisera raised against a partially purified preparation of the approximately equal to 60-kDa protective filarial antigen. These data relate the cloned antigen to a potentially protective antigen in lymphatic filariasis.
为便于对丝虫保护性抗原进行生化研究,从马来布鲁线虫成虫mRNA构建了一个λgt11 cDNA文库,并用能识别一组有限的约25、42、60和112 kDa丝虫抗原的兔血清进行筛选。先前已证明,约25 kDa和约60 kDa的抗原可诱导小鼠体内微丝蚴血症清除率提高。通过免疫反应检测到的一个154碱基对的克隆,用于通过杂交分离出一个近全长的1.8千碱基的cDNA克隆。核苷酸序列分析表明,该克隆来源于编码63 kDa抗原的mRNA。在免疫印迹实验中,用针对部分纯化的约60 kDa保护性丝虫抗原制备物产生的抗血清,识别出一种融合多肽,该融合多肽包含37 kDa的大肠杆菌色氨酸E蛋白(邻氨基苯甲酸合酶)和55 kDa的克隆蛋白。这些数据将克隆的抗原与淋巴丝虫病中一种潜在的保护性抗原联系起来。