The application discloses a beta-lactoglobulin
mutant, a preparation method and application, relates to the technical field of
genetic engineering, and the
amino acid sequence of the beta-lactoglobulin
mutant is shown as SEQ ID NO. 1; the preparation method is as follows: designing a
forward primer, a
reverse primer, a forward
mutant primer and a reverse mutant primer; using a wild-type beta-Lg
plasmid as a template, a mutant
DNA segment is obtained by using overlap extension PCR technology; after the mutant
DNA segment is connected with an empty vector, the obtained recombinant
plasmid is transformed into an E. coli
competent cell, cultured and identified; the recombinant
plasmid is transformed into the E. coli
competent cell, cultured and lysed; the
cell supernatant is collected, purified, and the beta-lactoglobulin mutant is obtained; the application improves the binding property of the beta-lactoglobulin with specific IgE antibodies, enhances the immunological activity of the beta-lactoglobulin
allergen, and enhances the sensitivity and specificity of the
allergen specific IgE
antibody detection result.