T Yuuki, Y Okumura, T Ando, H Yamakawa, M Suko, M Haida, M Dohi, H Okudaira
{"title":"Synthesis of biologically active recombinant Der f II.","authors":"T Yuuki, Y Okumura, T Ando, H Yamakawa, M Suko, M Haida, M Dohi, H Okudaira","doi":"10.1159/000235401","DOIUrl":null,"url":null,"abstract":"<p><p>A cDNA library corresponding to mite protein was screened employing anti-Der f II antibody. Two possible clones containing plasmids pFL1 and pFL11 were obtained. The two plasmids had insertions of about 500 basepairs. The DNA sequences of the two insertions were determined, from which the amino acid sequences were deduced. The amino acid sequence of the purified native Der f II protein could be determined to 45 residues from the N terminus. As a result of comparison, we concluded that the cDNAs prepared from live Dermatophagoides farinae mite corresponded to the mite allergen Der f II. The recombinant Der f II was biologically active.</p>","PeriodicalId":13810,"journal":{"name":"International archives of allergy and applied immunology","volume":"94 1-4","pages":"354-6"},"PeriodicalIF":0.0000,"publicationDate":"1991-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1159/000235401","citationCount":"9","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"International archives of allergy and applied immunology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1159/000235401","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 9
Abstract
A cDNA library corresponding to mite protein was screened employing anti-Der f II antibody. Two possible clones containing plasmids pFL1 and pFL11 were obtained. The two plasmids had insertions of about 500 basepairs. The DNA sequences of the two insertions were determined, from which the amino acid sequences were deduced. The amino acid sequence of the purified native Der f II protein could be determined to 45 residues from the N terminus. As a result of comparison, we concluded that the cDNAs prepared from live Dermatophagoides farinae mite corresponded to the mite allergen Der f II. The recombinant Der f II was biologically active.
利用抗derⅱ抗体筛选螨蛋白对应的cDNA文库。获得了含有质粒pFL1和pFL11的两个可能克隆。这两个质粒有大约500个碱基对的插入。测定了两个插入片段的DNA序列,并由此推导出氨基酸序列。纯化的天然Der f II蛋白的氨基酸序列可以从N端确定到45个残基。通过比较,我们得出结论,从活的粉蚧螨制备的cdna与螨变应原Der fⅱ相对应。重组蛋白具有生物活性。