کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
9283045 | 1225000 | 2005 | 12 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
The antigenic and catalytically active formamidase of Paracoccidioides brasiliensis: protein characterization, cDNA and gene cloning, heterologous expression and functional analysis of the recombinant protein
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کلمات کلیدی
CTABpCMBNonidet P-40nitrobluetetrazoliumTLCKNP40dithiotreitolParacoccidioidomycosisParacoccidioides brasiliensisFMD5-bromo-4-chloro-3-indolyl phosphateBCIPPMSFIEFNBTPCMGSTDTTIPTGPVPPVDFEDTA - اتیلن دی آمین تترا استیک اسید Ethylenediaminetetraacetic acid - اتیلینیدامین تتراستیک اسیدsodium dodecyl sulfate-polyacrylamide gel electrophoresis - الکتروفورز ژل دوده سولفات سدیم پلی آکریل آمیدSDS-PAGE - الکتروفورز ژل پلی آکریل آمیدisopropyl β-D-thiogalactopyranoside - ایزوپروپیل β-D-thiogalactopyranosideisoelectric focusing - تمرکز ذره ای الکتریکیpolyvinylidene difluoride - دی فلوئورید پلی وینیلیدینCatalytic activity - فعالیت کاتالیستیphenylmethylsulfonyl fluoride - فنیل متیل سولفونیل فلورایدYeast - مخمرMycelium - میسلیومpolyvinylpolypyrrolidone - پی ویوی پلیپیرلیدونhigh-performance liquid chromatography - کروماتوگرافی مایعی کاراHPLC - کروماتوگرافی مایعی کاراglutathione S-transferase - گلوتاتیون S-ترانسفراز
موضوعات مرتبط
علوم زیستی و بیوفناوری
ایمنی شناسی و میکروب شناسی
ایمونولوژی
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
Paracoccidioides brasiliensis is a well-characterized pathogen of humans. To identify proteins involved in the fungus-host interaction, P. brasiliensis yeast proteins were separated by liquid isoelectric focusing, and fractions were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis. Immunoreactive bands were detected with pooled sera of patients with P. brasiliensis infection. A protein species with a molecular mass of 45Â kDa was subsequently purified to homogeneity by preparative gel electrophoresis. The amino acid sequence of four endoproteinase Lys-C-digested peptides indicated that the protein was a formamidase (FMD) (E.C. 3.5.1.49) of P. brasiliensis. The complete cDNA and a genomic clone (Pbfmd) encoding the isolated FMD were isolated. An open reading frame predicted a 415-amino acid protein. The sequence contained each of the peptide sequences obtained from amino acid sequencing. The Pbfmd gene contained five exons interrupted by four introns. Northern and Southern blot analysis suggested that there is one copy of the gene in P. brasiliensis and that it is preferentially expressed in mycelium. The complete coding cDNA was expressed in Escherichia coli to produce a recombinant fusion protein with glutathione S-transferase (GST). The purified recombinant protein was recognized by sera of patients with proven paracoccidioidomycosis and not by sera of healthy individuals. The recombinant 45-kDa protein was shown to be catalytically active; FMD activity was detected in P. brasiliensis yeast and mycelium.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Microbes and Infection - Volume 7, Issue 1, January 2005, Pages 66-77
Journal: Microbes and Infection - Volume 7, Issue 1, January 2005, Pages 66-77
نویسندگان
Clayton L. Borges, Maristela Pereira, Maria S.S. Felipe, FabrÃcia P. de Faria, Francisco J. Gomez, George S. Jr., Célia M.A. Soares,