Nara F, Watanabe I, Serizawa N
Fermentation Research Laboratories, Sankyo Co., Ltd., Tokyo, Japan.
Curr Genet. 1993 Jan;23(1):28-32. doi: 10.1007/BF00336746.
We present here the first report of a transformation system developed for the filamentous, ML-236B (compactin)-producing fungus Penicillium citrinum. Hygromycin B-resistant colonies were obtained after treatment of protoplasts with a vector containing an Escherichia coli hygromycin B phosphotransferase gene fused to a 3-phosphoglycerate kinase promoter from Aspergillus nidulans. The transformation rate was 194 transformants per microgram circular DNA per 4 x 10(5) viable protoplasts under optimized transformation conditions. Transformation took place via the integration of plasmid DNA into the fungal chromosomal DNA. Most of the integration events appeared to produce tandemly iterated arrays of plasmid molecules at different sites in the chromosome. The transformed, drug-resistant, phenotype and the integrated plasmids were mitotically stable with or without selection in a majority of cases. The demonstration of such a transformation system is an essential first step in the application of recombinant DNA technology to strain improvement and for the production of novel ML-236B derivatives.
我们在此首次报道了一种为丝状、产ML - 236B（美伐他汀）的真菌桔青霉开发的转化系统。用含有与来自构巢曲霉的3 - 磷酸甘油酸激酶启动子融合的大肠杆菌潮霉素B磷酸转移酶基因的载体处理原生质体后，获得了潮霉素B抗性菌落。在优化的转化条件下，转化率为每微克环状DNA每4×10⁵个活原生质体产生194个转化体。转化是通过质粒DNA整合到真菌染色体DNA中实现的。大多数整合事件似乎在染色体的不同位点产生了串联重复的质粒分子阵列。在大多数情况下，无论有无选择压力，转化后的耐药表型和整合的质粒在有丝分裂过程中都是稳定的。这种转化系统的建立是将重组DNA技术应用于菌株改良以及生产新型ML - 236B衍生物的关键第一步。
