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非核糖体肽具抗生素、抗病毒、抗癌及免疫抑制剂等重要的药用价值。以长松萝的地衣型真菌为研究材料,通过同源克隆和Gene walking的方法获取长松萝地衣型真菌全长非核糖体肽合成酶基因(Us NRPS),并对得到的Us NRPS进行生物信息学分析、分子系统进化分析及基因表达分析。结果表明:在地衣型真菌中,非核糖体肽在长松萝中为首次发现,BLAST比对表明其为NRPS基因的A结构域,生物信息学分析显示Us NRPS蛋白是一种非分泌蛋白,位于细胞质基质中;分子进化分析显示Us NRPS与埃默森蓝状菌(Rasamsonia emersonii)的NRPS基因的A结构域蛋白聚为一类;RT-PCR分析表明:2%肌醇、10%甘露醇、2%山梨醇、2%葡萄糖、10%葡萄糖、0.2%谷氨酰胺、1%甘氨酸、0.2%丙氨酸能够诱导NRPS基因的轻微表达,2%、10%蔗糖和果糖均能诱导NRPS基因的强烈表达;0.2%、1%的天冬氨酸均抑制NRPS基因的诱导表达。
Non-ribosomal peptides with antibiotics, anti-virus, anti-cancer and immunosuppressive agents and other important medicinal value. Taking the lichen-type fungi of Pinus sylvestris as the research material, Us NRPS was obtained from the homologous clone and Gene walking method and the obtained Us NRPS Informatics analysis, molecular phylogenetic analysis and gene expression analysis. The results showed that non-ribosomal peptides were found for the first time in B. lugens. The BLAST alignment showed that it was the A domain of NRPS gene. Bioinformatics analysis showed that Us NRPS protein is a non-secreted protein, Located in the cytoplasm matrix; molecular evolution analysis showed that Us NRPS clustered with A domain proteins of NRPS gene of Rasamonia emersonii; RT-PCR analysis showed that 2% inositol, 10% mannitol , 2% sorbitol, 2% glucose, 10% glucose, 0.2% glutamine, 1% glycine, 0.2% alanine can induce slight expression of NRPS gene, NRPS gene can be induced by 2%, 10% sucrose and fructose 0.2% and 1% aspartate all inhibited the induced expression of NRPS gene.