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【目的】为了从放线菌发现新的药物先导化合物,研究了川滇4个地区的放线菌多样性及其生物活性。【方法】采集250份土样,用4种培养基分离放线菌;从中选择98株代表菌进行了初步分类鉴定;采用琼脂扩散法,检测了169株放线菌对4种细菌和7种真菌的抑菌活性;利用特异性引物扩增法,测定了它们产生的聚酮合酶(PKSI、PKSⅡ)基因、非核糖体多肽合成酶(NRPS)基因和多烯类化合物合成酶(CYP)基因。【结果】黄荆老林的放线菌有13个属,峨眉山、青城山仅5个属,九寨沟9个属,西双版纳达20个属;不同地区的放线菌具有抗菌活性的菌株平均约占10%;有27%-36%的菌株产生PKSI、II、NRPS、CPY化合物合成基因。【结论】在采集样品的地区中,人类干扰越少,放线菌的多样性越高。分离放线菌时,使用“极端”条件,虽然分离到的放线菌数量可能不多,但获得未知菌的比例较大。添加抑制剂可减少革兰氏阴性细菌和真菌,有利于分离放线菌。
【Objective】 In order to find new lead compounds from actinomycetes, the diversity of actinomycetes and their biological activity in four areas of Sichuan, Yunnan and Yunnan were studied. 【Method】 250 soil samples were collected and the actinomycetes were separated by four kinds of medium. 98 strains of bacteria were selected for preliminary classification and identification. The results of agar diffusion method showed that 169 strains of actinomycetes were sensitive to 4 kinds of bacteria and 7 species (PKSI, PKSⅡ), non-ribosomal polypeptide synthase (NRPS) gene and polyene synthase (CYP) synthase (CYP) gene. 【Result】 There were 13 genera of actinomycetes in Huangjing old forest, only 5 genera in Mount Emei and Qingcheng Mountain, 9 genera in Jiuzhaigou, and 20 genera in Xishuangbanna. Actinomycetes in different regions had an average of about 10% ; 27% -36% strains produce PKSI, II, NRPS, CPY compound synthetic genes. 【Conclusion】 In the area where samples were collected, the less human disturbance, the higher the diversity of actinomycetes. Actinomycete isolation, the use of “extreme ” conditions, although the number of isolated actinomycetes may be small, but the proportion of access to unknown bacteria larger. Adding inhibitors reduces Gram-negative bacteria and fungi and helps isolate actinomycetes.