RNAi技术降低头孢菌素C工业生产菌株中cefG基因的转录.pdfVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 2010 10 RNAi C cefG 龚桂花 刘艳 胡又佳 朱春宝 朱宝泉 ( , 200437) : RNA i是近年来新兴的一种通过抑制基因转录水平来 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 RNA i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 i cefG基因 头孢菌素 C 顶头孢霉 Downregulation of cefG Gene Transcription in an Industrial Strain of A cremon ium chrysog enum by RNA Interference Gong Guihua Liu Yan u Youjia ZhuChunbao Zhu Baoquan (Shanghai Institute of Pharmaceutical Industry, State K ey L aboratory of N ew D rug and P harmaceutical P rocess, Shanghai 200437) Abstrac:t RNA interference (RNA i) is a new technology to silence the expression of a target gene by suppressing its transcrip tional level. This paper usedRNA ito suppress the transcription of cefG gene in thebiosynthesis pathway ofCephalosporin C (CPC). A cefG gene silencing plasmid pYG273 bearing a phleomycin resistant gene and a transcriptional unit for doublestrand cefG RNA was constructed and transformed into an industrial strain ofA cremonium chry sogenum . The transcription levels of cefG gene in the transfor mantswere decreased by more than 80% at day 4 of fermentation, while CPC productionswere reduced by 34. 6% and 28. 8%, respec tively. K ey w ords: RNAi cefG gene Cephalosporin C A cremonium chry sogenum C(CPC) mRNA [4] RNA , , (N eurosp ora crassa ) , [ 5] (M ag nap orth