Chen Z, Song Y, Wen Y, Li J
Department of Microbiology, College of Biological Sciences, China Agricultural University, Beijing 100094, China.
Wei Sheng Wu Xue Bao. 2001 Aug;41(4):440-6.
Recombinant plasmid pCZ2(pKC1139::475 bp aveD) was used for aveD gene disruption in Streptomyces avermitilis 76-9. The plasmid was inserted into the chromosome by homogenous recombination between partial aveD gene in the plasmid and aveD in the chromosome. Disruptants were confirmed by Southern blotting. Shaking flask experiments and HPLC analysis showed that the disruptant produced only four components, which were C5-oxo-avermectin B1a, B1b, B2a, B2b as identified by UV, IR, NMR, and MS. This revealed that both aveD and aveF were not expressed in the disruptant. This is consistent with that aveD and aveF are in a transcription unit. This paper also provided a new genetic method to obtain C5-oxo-avermectin B-producing strain.
重组质粒pCZ2（pKC1139::475 bp aveD）用于阿维链霉菌76-9中aveD基因的破坏。该质粒通过质粒中的部分aveD基因与染色体中的aveD之间的同源重组插入到染色体中。通过Southern印迹法确认了破坏株。摇瓶实验和HPLC分析表明，该破坏株仅产生四种成分，经UV、IR、NMR和MS鉴定为C5-氧代阿维菌素B1a、B1b、B2a、B2b。这表明aveD和aveF在破坏株中均未表达。这与aveD和aveF在一个转录单元中是一致的。本文还提供了一种获得产生C5-氧代阿维菌素B菌株的新遗传方法。
