Laccases of white-rot fungi attract considerable attention due to their possible involvement in the transformation of a wide variety of phenolic compounds including the polymeric lignin and humic substances.In this research

a new simple method was developed for laccase cDNA cloning devoid RNA manipulation.After the fungi Ganoderma lucidum laccase gene that contain introns was cloned by the method of genome walking PCR.10 pair of primers were designed to amplify 10 exons.When these primers were designed

reward primer sequence of last exon must have 20 base homologous with forward primer sequence of next exon.Then

10 exons were amplied and recovered

respectively.Two exons in sequence were mixed as template to obtain 5 double-exon fragments.2 double-exon fragments were mixed as template to obtain large fragment exon1-4

and 3 double-exon fragments to obtain large fragment exon 5-10

respectively.At last

these two large fragments were mixed as template to obtain total cDNA sequence.Comparing with usual used

it is unnecessary to manipulate RNA and explore enzyme expression condition in this method.If this method combined with genomewakling PCR method can clone laccase gene and cDNA rapidly.