基于NRPS基因筛选与鉴定葡萄附生真菌
Isolation and identification based on NRPS functional gene of a fungal epiphytic strain from grapes
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摘要: 以非核糖体多肽合成酶（nonribosomal peptide synthetase,NRPS）基因为筛选靶点,从葡萄中筛选出含有NRPS基因的附生真菌,以期发现具有抗菌作用的环肽类化合物。采用平板划线技术、斜面分离纯化技术分离葡萄附生真菌,从中筛选出一株含有NRPS基因的附生真菌（编号PTLM-1）。在此基础上,构建系统发育进化树,结合形态学方法,将该菌株鉴定为腐皮镰刀菌Fusarium solani。经过NRPS系统发育进化树分析,发现该菌株有产生抗菌环肽类化合物——环孢菌素的潜力。通过对该菌的发酵产物进行电喷雾质谱分析,进一步证实了该预测结果。该研究为定向筛选产生环肽类化合物的菌株提供了新的研究方法与理论依据。Abstract: A fungal epiphytic with nonribosomal peptide synthetase( NRPS) gene was isolated from a root rot of grapes basing on nonribosomal peptide synthetase gene target- screening in order to find a cyclic peptide with antimicrobial activity.The fungal epiphytic with nonribosomal peptide synthetase( NRPS) gene named as PTLM- 1was isolated from grapes by streak plate and slant culture methods.On this basis,constructed a phylogenetic tree,then the species of the strain PTLM-1 was identified as F. solani combined with morphological observation. It was found that the strain had the potential to produce a cyclic peptide named cyclosporine after NRPS sequence analysising,which was further confirmed by the electrospray ionizsation mass spectrum( ESI- MS) analysis. This study provided new research methods and theoretical basis for target- screening strains which produced cyclic peptide.
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