Cloning and sequence analysis of laccase gene fragments from Climacocystis borealis

Two degenerate oligonucleotide primers were designed,coding for two conservative amino acid regions in fungi laccase protein family.A PCR amiplification was performed on Climacocystis borealis DNA template and a fragment of 1201 bp was obtained.By using the program of BLAST on NCBI GenBank database,the sequence presented a very high match with the laccase gene from other fungi and 99 searched sequences were all laccase genes.It is concluded that the cloned sequence is Climacocystis borealis laccase gene fragment. The sequence data have been submitted to the GenBank database under accession number AY333124. According to the ClustalW comparison analysis with Coriolopsis gallica(laccase lcc 1)cds,six introns are included and the sequence codes 286 amino acids after excluding the introns.Through ClustalW analysis with other fungi laccase genes,it is found that the Climacocystis borealis laccase amino acid fragment shares considerable homology of above 51%.