Bull J H, Smith D J, Turner G
Department of Microbiology, Medical School, Bristol, UK.
Curr Genet. 1988 May;13(5):377-82. doi: 10.1007/BF00365658.
We have cloned a mutant oligomycin resistance allele of the mitochondrial ATP synthase subunit 9 gene from the filamentous fungus Penicillium chrysogenum. The gene was isolated using the equivalent gene from Aspergillus nidulans as a hybridisation probe. Using the cloned gene it is possible to select for oligomycin resistance in P. chrysogenum transformation experiments. This transformation system was used to introduce further copies of the P. chrysogenum isopenicillin N synthetase gene, which were stably maintained without selection. An assessment of the frequency with which homologous integration occurs was also made. With this system, it should prove possible to transform any strain of P. chrysogenum.
我们从丝状真菌产黄青霉中克隆了线粒体ATP合酶亚基9基因的一个突变型寡霉素抗性等位基因。该基因是用构巢曲霉的等效基因作为杂交探针分离得到的。利用克隆的基因,在产黄青霉转化实验中可以筛选出寡霉素抗性。这个转化系统被用于导入产黄青霉异青霉素N合成酶基因的更多拷贝,这些拷贝在无选择压力的情况下能稳定维持。还对同源整合发生的频率进行了评估。利用这个系统,应该能够转化产黄青霉的任何菌株。