Isolation and cloning of manganese peroxidase (mnp) gene from oyster mushroom

Different enzymes in oyster mushroom (pleurotus oatreatus), can degradate the lignin compounds from the beginning of mycelium growth up to the end of fruiting period in the environment of compost and straw. Wood, plant residue and most of plant wastes in the nature are called lignocelluloses compounds. Lignocelluloses is mainly composed of cellulose, hemicelluloses and lignin. Magnesium peroxides enzyme (EC:1:11:1:13) is one the most common peroxides destroying lignin which produced by most wood decomposing mushrooms as well as many compost decomposing mushrooms. In order to cloning of mnp gene, RNA was extracted from oyster edible mushroom (P.ostreatus var.florida) and cDNA was synthesized and then the primers was designed based on the sequence of the mnp gene using Primer Premier (V.5.0) software and was amplified by PCR. First, the gene was inserted to pTG-19T plasmid and was confirmed using sequencing. In continue, mnp gene was cloned in to the p13H88 plasmid. Then it was transferred to Ecoli (DH5a) using ice-melt method and its presence was confirmed by enzymatic digestion. The results showed that mnp gene is cloned in p13H88 plasmid and the recombinant plasmid was named as p13H88-FM.