Cullen D, Leong S A, Wilson L J, Henner D J
Forest Products Laboratory, USDA, Madison, WI 53705.
Gene. 1987;57(1):21-6. doi: 10.1016/0378-1119(87)90172-7.
Aspergillus nidulans strain G191 was transformed to hygromycin resistance using plasmid pDH25, which contains the bacterial hygromycin B phosphotransferase gene (hph) fused to promoter elements of the A. nidulans trpC gene. Southern hybridizations of transformants revealed multiple, integrated copies of the vector. A pleiotropic effect conferring increased hygromycin B sensitivity was found to be associated with the A. nidulans pyrG89 allele. Plasmid pDH25 features a ClaI site immediately preceding the hph start codon thus permitting convenient replacement of the trpC sequences with other eukaryotic promoters.
使用质粒pDH25将构巢曲霉G191菌株转化为潮霉素抗性,该质粒含有与构巢曲霉trpC基因启动子元件融合的细菌潮霉素B磷酸转移酶基因(hph)。转化体的Southern杂交显示载体有多个整合拷贝。发现构巢曲霉pyrG89等位基因与赋予潮霉素B敏感性增加的多效性效应有关。质粒pDH25在hph起始密码子之前紧邻一个ClaI位点,因此允许用其他真核启动子方便地替换trpC序列。