Binninger D M, Skrzynia C, Pukkila P J, Casselton L A
EMBO J. 1987 Apr;6(4):835-40. doi: 10.1002/j.1460-2075.1987.tb04828.x.
We have developed a simple and efficient transformation system for the agaric fungus, Coprinus cinereus. Protoplasts were prepared from asexual spores that harbor one or two mutations in the structural gene for tryptophan synthetase. The protoplasts can be stably transformed using the cloned Coprinus gene at a frequency of 1 in 10(4) viable protoplasts. A variety of molecular events accompanies the formation of stable transformants, including insertion of the transforming DNA at the homologous locus. The transforming DNA is stable through cell division, mating, fruiting body formation, and meiosis.
我们已经为灰盖鬼伞这种伞菌开发了一种简单高效的转化系统。原生质体是从色氨酸合成酶结构基因中携带一到两个突变的无性孢子制备而来的。这些原生质体可以用克隆的灰盖鬼伞基因以每10⁴个活原生质体中有1个的频率进行稳定转化。稳定转化体的形成伴随着多种分子事件,包括转化DNA在同源位点的插入。转化DNA在细胞分裂、交配、子实体形成和减数分裂过程中都是稳定的。