人参大片段DNA(100kb)转化灵芝的研究

Transformation of large DNA fragment of Panax ginseng into Ganoderma lucidum

为探讨人参大片段DNA转化灵芝的可能性,通过电击法将双元细菌人工染色体(BIBAC)载体上的100kb人参大片段DNA转化到灵芝原生质体内。研究发现,在电极间距为4mm,电压强度为240V时,将5μL的人参大片段DNA转化到75μL的灵芝原生质体,在选择培养基上获得了具有再生能力的转化子。根据克隆载体两侧的序列设计两对引物,对转化子进行PCR分析。试验结果表明人参大片段DNA已经转化到灵芝的基因组中。

A 100kb DNA fragment of Panax ginseng cloned in a binary bacterial artificial chromosome （BIBAC） vector was transformed into protoplasts of Ganoderma lucidum by electroporation. The results showed that a number of regenerative transformants were obtained on the selective medium when 5μL of the Panax ginseng large DNA fragment solution was transformed into 75μL of the Ganoderma lucidum protoplasts using 4mm spaced electrodes at 240 volts. The transformants were analyzed by PCR using two pairs of primers designed from two flanking sequence of the vector cloning site. The results indicated that the ginseng large DNA fragment had been transformed into the Ganoderma lucidum genome.