The invention specifically discloses a preparation method and application of a recombination ryegrass
pollen allergen and a
mutant thereof. The recombination ryegrass
pollen allergen disclosed by the invention is a
mutant which exists in a non-natural manner and is derived by an
allergen naturally existing; and the
amino acid sequence of the recombination ryegrass
pollen allergen is shown as SEQIDNO.1. According to the preparation method and application disclosed by the invention, a coding
gene of the recombination ryegrass
pollen allergen is artificially synthesized by optimizing a codon by using a
genetic engineering technology; an antigenic
epitope of the coding
gene is analyzed by adopting
bioinformatics software; and the directional displacement for one or more loci with high allergen is realized by site-specific
mutagenesis, and thus the allergic endogenous property of the loci is reduced and the
mutant of the recombination ryegrass
pollen allergen with low allergic endogenous property is constructed. The
protein expression is carried out by artificial control, and thus the recombination ryegrass
pollen allergen with high purity and a
mutant protein of the recombination ryegrass pollen allergen are obtained. Compared with natural allergen, the recombination ryegrass pollen allergen has the characteristic that the combining capacity between the recombination
protein mutant and specific IgE (
Immunoglobulin E) is remarkably reduced; and the recombination ryegrass pollen allergen can be safely applied to diagnosis and treatment of allergic diseases even the judgment for the levels of other allergic endogenous properties.