Hilber U W, Bodmer M, Smith F D, Köller W
Department of Plant Pathology, Swiss Federal Research Station, Wädenswil.
Curr Genet. 1994 Feb;25(2):124-7. doi: 10.1007/BF00309537.
Botryotinia fuckeliana, the causal agent of grey mould, was biolistically transformed to hygromycin B resistance using a plasmid (pOHT) containing a bacterial hygromycin phosphotransferase gene fused to regulatory sequences from Aspergillus nidulans. Multiple copies of the plasmid, precipitated onto tungsten particles, were delivered into the conidia by a helium-driven gene gun. Southern analysis showed that the plasmid was integrated into the fungal genome at one single locus. After five subsequent transfers on selective medium, all transformants were mitotically stable. When propagated on non-selective medium, four out of eight transformants retained their resistance to hygromycin B. Southern analysis of the fifth generation of transformants showed that no genetic rearrangements occurred during vegetative growth of stable transformants.
灰霉病的病原菌富克尔核盘菌（Botryotinia fuckeliana）通过使用一种质粒（pOHT）进行生物弹射击转化，使其对潮霉素B产生抗性，该质粒含有一个与来自构巢曲霉调控序列融合的细菌潮霉素磷酸转移酶基因。沉淀在钨颗粒上的多个质粒拷贝通过氦驱动基因枪导入分生孢子中。Southern分析表明，该质粒在单个位点整合到真菌基因组中。在选择性培养基上连续传代五次后，所有转化体在有丝分裂过程中都是稳定的。当在非选择性培养基上传代时，八个转化体中有四个保持了对潮霉素B的抗性。对第五代转化体的Southern分析表明，稳定转化体营养生长期间未发生基因重排。
