非核糖体肽合成酶缩合结构域基因片段克隆

从大连渤海海域筛选出1株放线菌L1,结合形态观察、生理生化实验和16S rDNA分子鉴定,确定L1属于链霉菌属球孢链霉菌(Streptomyces globisporus)。根据GenBank发布的非核糖体肽合成酶(NRPS)序列设计引物,从放线菌L1的基因组DNA中扩增获得NRPS基因片段。测序结果及比对分析表明该片段属于NRPS缩合结构域部分序列。三维建模显示其结构呈V型,包含缩合结构域核心序列,与数据库已知结构相一致,可以推断该克隆片段为NRPS缩合结构域基因片段,为后续深入研究缩合结构域特异性与相关NRPS功能提供基础。

Cloning of nonribosomal peptide synthetase gene condensation domain fragment

One actinomycetes strain L1 was isolated from seaweed and algae in Dalian sea area.Combined with morphology observation,physiological and biochemical experiments and 16S rDNA identification,L1 belonged to Streptonyces globisporus.One pair of primers was designed and a 400 base pair fragment was amplified from L1 genomic DNA.The results revealed that the cloned fragment belonged to part of NRPS condensation domain sequence.Three dimensional modeling of the cloned fragment showed that its structure had type V morphology,and the core sequence of the condensation domain was consistent with the known structure in database.It could inferred that the cloned fragment was NRPS condensation domain gene fragment,which would provide the basis for further study of condensation domain specificity and related NRPS function.