论文部分内容阅读
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。
In order to explore the possibility of transforming ginseng with large fragment DNA of ginseng, 100kb large DNA fragment of ginseng in BIBAC vector was transformed into Ganoderma protoplast by electroporation. The results showed that when the electrode spacing was 4mm and the voltage was 240V, 5μL of large ginseng DNA was transformed into 75μL of protoplast of Ganoderma lucidum, and the transformant with regenerative ability was obtained on selective medium. Two pairs of primers were designed according to the sequences on both sides of the cloning vector, and the transformants were subjected to PCR analysis. The test results show that large fragments of ginseng DNA has been transformed into the genome of Ganoderma lucidum.