Tilburn J, Scazzocchio C, Taylor G G, Zabicky-Zissman J H, Lockington R A, Davies R W
Gene. 1983 Dec;26(2-3):205-21. doi: 10.1016/0378-1119(83)90191-9.
DNA-mediated genetic transformation of Aspergillus nidulans has been achieved by incubating protoplasts from a strain of A. nidulans carrying a deletion in the acetamidase structural gene with DNA of derivatives of plasmid pBR322 containing the cloned structural gene for acetamidase [Hynes et al., Mol. Cell. Biol. 3 (1983) 1430-1439; p3SR2] in the presence of polyethylene glycol and CaCl2. The highest frequency obtained was 25 transformants per microgram of DNA. No enhancement of the transformation frequency was observed when DNAs of plasmids carrying either a fragment of the A. nidulans ribosomal repeat (p3SR2rr) or a fragment containing a possible A. nidulans mitochondrial origin of replication (p3SR2mo) in addition to the acetamidase gene were used. Both pBR322 and acetamidase gene sequences become integrated into the genome of A. nidulans in transformant strains. Integration events into the residual sequences adjacent to the deletion in the acetamidase gene, and probably (for p3SR2rr and p3SR2mo) into the ribosomal repeat unit are described.
通过将携带乙酰胺酶结构基因缺失的构巢曲霉菌株的原生质体与含有克隆的乙酰胺酶结构基因的质粒pBR322衍生物的DNA [海因斯等人，《分子细胞生物学》3 (1983) 1430 - 1439；p3SR2] 在聚乙二醇和氯化钙存在的情况下孵育，已实现了构巢曲霉的DNA介导的遗传转化。获得的最高频率是每微克DNA有25个转化体。当使用除乙酰胺酶基因外还携带构巢曲霉核糖体重复片段（p3SR2rr）或含有可能的构巢曲霉线粒体复制起点片段（p3SR2mo）的质粒DNA时，未观察到转化频率的提高。在转化菌株中，pBR322和乙酰胺酶基因序列都整合到了构巢曲霉的基因组中。描述了整合到乙酰胺酶基因缺失旁的残留序列中，以及可能（对于p3SR2rr和p3SR2mo）整合到核糖体重复单元中的整合事件。
