Development of vector applicable to basidiomycetes using lignin-degrading gene as the marker.

本研究旨在分离担子菌寄主-载体系统构建所必需的标记基因，通过聚合酶链式反应从平菇基因组DNA中扩增出与锰(II)过氧化物酶基因相对应的DNA标记，并以此为探针，通过Southern杂交从平菇基因组DNA中分离出锰(II)过氧化物酶基因。测定了杂交法获得的克隆的核苷酸序列。对获得的克隆进行二次筛选。从平菇菌丝体中分离到两个线粒体质粒(PO-1和PO-2)，利用RT-PCR技术对平菇菌丝体中MNP活性前的转录进行了鉴定。将约55%的空洞DNA克隆到大肠杆菌载体中。

This research aims to isolate the marker gene which are indispensable for the construction of host-veetor system of basidiomycetes.DNA flagments which were thought to correspond to the Mn (II) peroxidase (MnP) gene were obtained from genomic DNA of Pleurotus ostreatus by PCR.These flagments were further used as the probes to isolate the DNA of MnP gene by Southern hybridization from the genomic DNA of P.ostreatus. Nucleotide sequence obtained from the clones by the hybridization were detemined. Secondary screening was carried out on the clones obtained. Transcription of mRNA isolated from the mycelia of P.ostreatus just before the activity of MnP was certificated by using RT-PCR technique.Two lenear mitochondrial plasmids (PO-1 and-2) were isolated from the mycelia of P.ostreatus. About 55 % regeon of the hole DNA was cloned in to the vectors of Esherichia coli.