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Cloning, sequence analysis, and expression of cDNA coding for the major house dust mite allergen, Der f 1, in Escherichia coli BJMBR
Cui,Y.; Zhou,P.; Peng,J.; Peng,M.; Zhou,Y.; Lin,Y.; Liu,L..
Our objective was to clone, express and characterize adult Dermatophagoides farinae group 1 (Der f 1) allergens to further produce recombinant allergens for future clinical applications in order to eliminate side reactions from crude extracts of mites. Based on GenBank data, we designed primers and amplified the cDNA fragment coding for Der f 1 by nested-PCR. After purification and recovery, the cDNA fragment was cloned into the pMD19-T vector. The fragment was then sequenced, subcloned into the plasmid pET28a(+), expressed in Escherichia coli BL21 and identified by Western blotting. The cDNA coding for Der f 1 was cloned, sequenced and expressed successfully. Sequence analysis showed the presence of an open reading frame containing 966 bp that encodes a...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Dermatophagoides farinae; Der f 1; Recombinant allergens; Cloning; Expression; Bioinformatics.
Ano: 2008 URL:
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