Mach R L, Schindler M, Kubicek C P
Abteilung für Mikrobielle Biochemie, TU Wien, Austria.
Curr Genet. 1994 Jun;25(6):567-70. doi: 10.1007/BF00351679.
Trichoderma reesei was transformed to hygromycin B resistance using a novel vector, which contains the E. coli hygromycin B phosphotransferase gene (hph) fused between promoter and terminator elements of the homologous Trichoderma pki1 (coding for pyruvate kinase) and cbh2 (coding for cellobiohydrolase II) genes, respectively. Transformation frequencies of over 1,800--2,500 transformants/micrograms DNA were obtained, which is a 15--20-fold increase over that with pAN7-1, which contains hph between A. nidulans expression signals. Mitotically-stable transformants contained the hph gene and the regulatory sequences of the pki1 promoter and the cbh2 terminator integrated into the genome. Evidence for preferentially ectopic integration is given.
里氏木霉通过一种新型载体转化为对潮霉素B具有抗性，该载体包含大肠杆菌潮霉素B磷酸转移酶基因（hph），分别融合在同源里氏木霉pki1（编码丙酮酸激酶）和cbh2（编码纤维二糖水解酶II）基因的启动子和终止子元件之间。获得了超过1800 - 2500个转化子/微克DNA的转化频率，这比含有构巢曲霉表达信号之间的hph的pAN7 - 1提高了15 - 20倍。有丝分裂稳定的转化子含有整合到基因组中的hph基因以及pki1启动子和cbh2终止子的调控序列。给出了优先异位整合的证据。
