Molecular cloning and expression analysis of a F-type lectin gene from Japanese sea perch (Lateolabrax japonicus).

摘要

The techniques of homology cloning and anchored PCR were used to clone the fucose-binding lectin (F-type lectin) gene from Japanese sea perch (Lateolabrax Japonicus). The full-length cDNA of sea perch F-lectin (JspFL) contained a 5' untranslated region (UTR) of 39[NON-BREAKING SPACE]bp, an ORF of 933[NON-BREAKING SPACE]bp encoding a polypeptide of 310 amino acids with an estimated molecular mass of 10.82[NON-BREAKING SPACE]kDa and a 3' UTR of 332[NON-BREAKING SPACE]bp. The searches for nucleotides and protein sequence similarities with BLAST analysis indicated that the deduced amino acid sequence of JspFL was homological to the Fucose-binding lectin in other fish species. In the JspFL deduced amino acid sequence, two tandem domains that exhibit the eel carbohydrate-recognition sequence motif were found. The temporal expressions of gene in the different tissues were measured by real-time PCR. And the mRNA expressions of the gene were constitutively expressed in tissues including spleen, head-kidney, liver, gill, and heart. The JspFL expression in spleen was different during the stimulated time point, 2[NON-BREAKING SPACE]h later the expression level became up-regulated, and 6[NON-BREAKING SPACE]h later the expression level became down-regulated. The result indicated that JspFL was constitutive and inducible expressed and could play a critical role in the host-pathogen interaction.