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Expression, Characterization and Antigenicity Evaluation and 3D Structural Modeling of Peanut Allergen iso-Ara h 3
Author: ZhongHaiFeng
Tutor: MaSanMei
School: Jinan University
Course: Biochemistry and Molecular Biology
Keywords: Peanut allergens Cloning and expression Immune identification Antigenicity Three-dimensional structure
CLC: S565.2
Type: Master's thesis
Year: 2009
Downloads: 47
Quote: 0
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Abstract
Food allergy is considered a more serious nutritional problems , has aroused widespread concern . Peanut ( Arachis hypogaea.L ) is a nutritious protein resources . At the same time , in the world, FAO ( FAO , 1995 ) reported eight categories of food allergens , peanut allergens is important . Peanut allergy with a low dose sensitization, high mortality and life along with other characteristics, and therefore also been extensively studied . In this study, Touch down PCR technology , from peanut cDNA clone the peanut allergens iso - Ara h 3 gene , the results obtained two clones were named of PN32 - 8 and PN32 - 20 . Sequencing and alignment analysis , initially identified as the - clone basis because the target gene , and the target gene into the expression vector pET-44a was transformed into E. coli induced expression . Target gene expression by SDS-PAGE confirmed the 58 kDa protein , Western blot technology while taking advantage of the recombinant protein antigen identification . The results showed that recombinant iso - Ara h 3 protein serum recognition rate of 12.5% ??, and peanut major allergen the original Ara h 3 is a weak compared to the allergenic protein . Finally, bioinformatics antigenic analysis of the recombinant protein , and in the three-dimensional structure of the successful implementation of single amino acids virtual transformation .
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CLC: > Agricultural Sciences > Crop > Economic crops > Oil crops > Peanut
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