Tang G, Tang W, Zhong L, Yang K
Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
Chin J Biotechnol. 1996;12(2):131-6.
A 850 bp fragment of the 5' flanking region of the glucoamylase gene (glaA) was synthesized from A. niger T21 genome using PCR. The function detection vector was constructed by fusing this fragment to E. coli hygromycin B phosphotransferase gene (hph) and was used to transform A. niger. The high hygromycin resistance transformants thus obtained verified that the synthesized fragment functioned as a promoter in filamentous fungi. Southern blot analysis showed the hph gene has been integrated into genomic DNA of A. niger transformant.
使用聚合酶链反应(PCR)从黑曲霉T21基因组中合成了葡糖淀粉酶基因(glaA)5'侧翼区域的一个850 bp片段。通过将该片段与大肠杆菌潮霉素B磷酸转移酶基因(hph)融合构建功能检测载体,并用于转化黑曲霉。由此获得的高潮霉素抗性转化体证实了合成片段在丝状真菌中起启动子的作用。Southern杂交分析表明hph基因已整合到黑曲霉转化体的基因组DNA中。