Document Detail


Cloning, expression, and immunological characterization of recombinant Lolium perenne allergen Lol p II.
MedLine Citation:
PMID:  7691817     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The molecular cloning of the cDNA encoding for an isoallergenic form of Lol p II, a major rye grass (Lolium perenne) pollen allergen, was performed by polymerase chain reaction amplification on mRNA extracted from pollen. The amino acid sequence derived from the cDNA was truncated by 4 and 5 residues at the NH2- and COOH-terminal ends, respectively, and differed only in one position from that previously reported. This cDNA was expressed in Escherichia coli by fusion to the carboxyl terminus of the human ferritin H-chain. The molecule was produced in high yields as a soluble protein and was easily purified. The protein retains the multimeric quaternary structure of ferritin, and it exposes on the surface the allergenic moiety, which can be recognized in Western blotting and in enzyme-linked immunosorbent assay experiments by specific IgE from allergic patients. The recombinant allergen was used to analyze the sera of 26 patients allergic to L. perenne compared with control sera. The results were in good agreement with the values obtained with the radioallergosorbent test assay. In addition, histamine release experiments in whole blood from an allergic patient and skin prick tests showed that the recombinant allergen retains some of the biological properties of the natural compound. These findings indicate that the availability of homogeneous recombinant allergens may be useful for the development of more specific diagnostic and therapeutic procedures. Moreover, this expression system may be of more general interest for producing large amounts of soluble protein domains in E. coli.
Authors:
A Sidoli; E Tamborini; I Giuntini; S Levi; G Volonté; C Paini; C De Lalla; A G Siccardi; F E Baralle; S Galliani
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  The Journal of biological chemistry     Volume:  268     ISSN:  0021-9258     ISO Abbreviation:  J. Biol. Chem.     Publication Date:  1993 Oct 
Date Detail:
Created Date:  1993-11-18     Completed Date:  1993-11-18     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  2985121R     Medline TA:  J Biol Chem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  21819-25     Citation Subset:  IM    
Affiliation:
Dipartimento di Ricerca Biologica e Tecnologica, Istituto Scientifico H San Raffaele, Milano, Italy.
Data Bank Information
Bank Name/Acc. No.:
GENBANK/X73363
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MeSH Terms
Descriptor/Qualifier:
Allergens / biosynthesis,  genetics*,  immunology
Amino Acid Sequence
Base Sequence
Cloning, Molecular
DNA, Complementary
Escherichia coli
Histamine Release
Humans
Lolium / immunology
Molecular Sequence Data
Plant Proteins / biosynthesis,  genetics*,  immunology
Recombinant Proteins / biosynthesis,  genetics,  immunology
Sequence Homology, Amino Acid
Skin Tests
Chemical
Reg. No./Substance:
0/Allergens; 0/DNA, Complementary; 0/Lol p II protein, Lolium perenne; 0/Plant Proteins; 0/Recombinant Proteins

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