Abstract
On the basis of eight full-length cDNA base sequences of laccase from Trametes versicolor (T. versicolor) registered in the sequence data base of National Center of Biotechnology Information (NCBI), laccases from T. versicolor were classified into four groups (laccase 1-4) according to the sequence homology over 95%. Four primer sets for the cloning of each of four laccase genes were prepared, respectively, and then the cloning of full-length complementary DNA (cDNA) for each of four genes was performed using reverse transcriptase-polymerase chain reaction (RT-PCR) method with the total RNA from T. versicolor as a template and each primer set. As a result, each full-length cDNA of four genes was obtained, and that the sequence homology among cDNAs of four genes was not more than ca. 75%. The synthesis of laccase protein for each of four laccase genes was confirmed using the in vitro expression system with wheat germ, and each protein having molecular weight of ca. 56, 000 that is the same as the calculated mass in accordance with the predicted amino acid sequence was detected on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).
