| 摘要 |
PURPOSE:To provide a novel protoplast hybrid cell having the drug component (antitumor active component) of Fomes japonicus and the proliferability and edibility of Cortinellus shiitake, and obtained by the cell fusion of Fomes japonicus and Cortinellus shiitake. CONSTITUTION:The pieces of the tissue of fruit body of Fomes japonicus and Cortinellus shiitake are implanted aseptically in Hamada's culture base for Armillaria edodes to effect the growth of mycelia. The mycelia re proliferated, transplanted in liquid media, subcultured, and treated with an enzyme liquid containing a combination of cellulase, dolicellase and thymoliase at 25 deg.C for 3hr to separate the protoplast of the mycelia of respective fungi. The separated protoplast is filtered, centrifuged at 700xg, and washed by repeating the dispersion in saline water and the centrifugal separation. The residue is dispersed in 30% PEG solution at 30 deg.C and incubated for 10min. The suspension is added with saline water and centrifuged, and the supernatant liquid is discarded. The objective protoplast hybrid cell can be obtained by the above operation. |
