Irie T, Honda Y, Hirano T, Sato T, Enei H, Watanabe T, Kuwahara M
Wood Research Institute, Kyoto University, Uji, Japan.
Appl Microbiol Biotechnol. 2001 Sep;56(5-6):707-9. doi: 10.1007/s002530100651.
It was reported that Pleurotus ostreatus was transformed unstably using recombinant plasmids containing a hygromycin B phosphotransferase gene (hph) under the control of Aspergillus nidulans expression signals, and that the plasmids were maintained extrachromosomally in the transformants. Here we report a stable and integrative transformation of the fungus to hygromycin B resistance, using a recombinant hph fused with Lentinus edodes glyceraldehyde-3-phosphate dehydrogenase expression signals. Restriction-enzyme-mediated integration (REMI) was also tried and increased the transformation efficiency about ten-fold.
据报道,利用含有潮霉素B磷酸转移酶基因(hph)且受构巢曲霉表达信号控制的重组质粒对糙皮侧耳进行转化时,转化不稳定,并且这些质粒在转化子中以染色体外形式维持。在此,我们报道了使用与香菇3-磷酸甘油醛脱氢酶表达信号融合的重组hph对该真菌进行稳定的整合转化,使其获得潮霉素B抗性。我们还尝试了限制酶介导的整合(REMI),其使转化效率提高了约十倍。