Obtaining an expression vector containing the nucleotide sequence of the gene encoding Bet v 1 – the major allergen of birch pollen

Abstract

An expression vector containing the gene encoding the most common isoform Bet v 1.0101 of the Bet v 1 protein, the major birch pollen allergen, was created for a subsequent expression in the prokaryotic system of Escherichia coli. Total RNA from birch pollen collected in Belarus was used as a matrix. The expression vector pJC40-Веt v 1 was obtained using molecular-genetic methods: cloning, ligation, transformation. The specificity of the cloned fragment was confirmed by sequencing. During the study, 14 rare Escherichia coli codons were identified in the coding part of the cloned gene. The triplets were evenly arranged in the nucleotide sequence; codon clustering was observed only in two cases. The total percentage of rare triplets (8.75 %) and the calculated value of the codon adaptation level (0.57) allow us to predict a sufficiently efficient expression of the studied gene. The data obtained will be used in the synthesis of the recombinant polypeptide Bet v 1.0101.