Amplification and Expression of a Lectin Gene from Agaricus bisporus AS2796

  Objective  A lectin gene from Agaricus bisporus AS2796 was isolated to study the functions of the agglutinin protein in mushrooms.

  Method  Based upon the reported sequences, a primer was designed to amplify the lectin gene with the total DNA as templates. A 432 bp PCR product was purified and connected to the prokaryotic expression vector pET-28a carrying the His label, then transformed into E. coli BL21 (DE3). The IPTG-induced expression product was subsequently purified by affinity chromatography and identified by Western blot.

  Result  The amplified lectin gene was 432 bp with correctly constructed recombinant expression plasmid, pET-lectin. The recombinant agglutinin protein was about 18 kDa and more than 95% pure after purification,

  Conclusion  For the first time, a lectin gene was successfully isolated from A. bisporus and amplified. The agglutinin protein obtained could be used to further study its bioactivity and biosynthesis.