摘要
nsdA(Negative regulator of Streptomyces differentiation)基因是天蓝色链霉菌中发现的与抗生素合成相关的负调控基因。根据天蓝色链霉菌A3(2)的序列设计引物扩增玫瑰黄链霉菌Men-myco-93-63菌株中的该基因,经测序和序列分析表明,由保守序列引物nsdA-2R和nsdA-2L克隆到的Men-myco-93-63菌株中的该基因800 bp片段与已知nsdA基因核苷酸保守区域序列同源性达到99%,由全长序列扩增引物nsdA WZ-R和nsdA WZ-L克隆到的该基因包含一个完整的开放阅读框,共编码369个氨基酸,与变铅青链霉菌和天蓝色链霉菌A3(2)中的nsdA基因的核苷酸序列同源性达到100%,氨基酸序列同源性达到99%。该全长基因命名为nsdAmgh,其在玫瑰黄链霉菌Men-myco-93-63中的发现为阻断该负调控基因以期获得抗生素高产工程菌株提供了重要依据。
nsdA (Negative regulator of Streptomyces differentiation )is globally negative regulator gene first found in Streptomyces coelicolor. In this study, nsdA gene of Streptomyces roseoflavus strains Men-myco-93-63 was amplified using the primer sequences designed from. Streptomyces coelicolor A3 (2). After sequencing, Blastn, and Blastx, it showed that the cloned fragment from Streptomyces roseoflavus Men-myco-93-63 with the primer of nsdA-2R and nsdA-2L compairing with reported nsdA conservative nucleotide sequence homology on the region reached 99 %. The full-length gene sequence of Men-myco-93-63 nsdA gene amplification with primers of nsdA WZ-R and nsdA WZ-L had a complete open reading frame encoding 369 amino acids, with the Streptomyces lividans and Streptomyces coelicolor A3 (2)nsdA gene at nucleotide sequence 100%, and amino acid sequence homology up to 99% . The full-length nsdA gene of Streptomyces roseoflavus Men-myco-93-63 was named nsdAmgh, which will promote the obtaintion of high-yielding antibiotic strains of Streptomyces roseoflavus Men-myco-93-63 through interrupting the negative regulator.
出处
《华北农学报》
CSCD
北大核心
2009年第5期77-80,共4页
Acta Agriculturae Boreali-Sinica
基金
国家自然科学基金项目(30800734)
河北省自然科学基金项目(C200900598)
国家高技术研究与发展计划“863”项目(2006AA10A211)