RNAi技术降低头孢菌素C工业生产菌株中cefG基因的转录

Down-regulation of cefG Gene Transcription in an Industrial Strain of Acremonium chrysogenum by RNA Interference

RNAi是近年来新兴的一种通过抑制基因转录水平来实现特定基因表达沉默的技术。本研究尝试在顶头孢霉工业生产菌株中利用RNAi技术沉默cefG基因的转录,构建了一个含可形成cefG双链RNA转录单元的质粒,并将其导入头孢菌素C工业生产菌株中。结果发现其中两个转化子的cefG基因转录水平在发酵第4天较出发菌株降低了80%以上,而它们的头孢菌素C发酵水平较出发菌株则分别降低了34.6%和28.8%。

RNA interference （RNAi） is a new technology to silence the expression of a target gene by suppressing its transcriptional level. This paper used RNAi to suppress the transcription of cefG gene in the biosynthesis pathway of Cephalosporin C （CPC）. A cefG gene silencing plasmid pYG273 bearing a phleomyein resistant gene and a transcriptional unit for double-strand eefG RNA was constructed and transformed into an industrial strain of Acremonium chrysogenum. The transeription levels of cefG gene in the transformants were decreased by more than 80% at day 4 of fermentation,while CPC productions were reduced by 34.6% and 28.8% ,respeetively.