Abstract: Perlucin was originally isolated from nacre of abalone inner shell and improved to promote calcium carbonate precipitation at ambient conditions, nucleate the calcium carbonate crystallization and modify the morphology of calcium carbonate crystals. The open reading frame（ORF）of the perlucin gene was amplified from the mantle cDNA of the freshwater pearl mussel, Hyriopsis cumingii. The ORF from this species was ligated into the expression plasmid pET-28a. Following transfer of the recombinant plasmid into host bacteria, Escherichia coli BL21（DE3）cells, the recombinant pET-28a-Hcperlucin protein was expressed by induction with isopropyl-β-thiogalactopyranoside（IPTG）. SDS-PAGE analysis showed that inducing the cells at 37℃ in 0.1 mmol/L of IPTG for 6 h was the optimal conditions for expression of the recombinant pET-28a-Hcperlucin protein. Inclusion body was the main existence form of the recombinant protein. The expression products were detected by SDS-PAGE and Western blot. The results showed that the recombinant expression plasmid of pET-28a-Hcperlucin, expressing a 21 kD protein that could be recognized by mouse anti-His-tag Mab, was successfully constructed.

Key words: Hyriopsis cumingii, Perlucin, Prokaryotic expression Western blot analysis