目的:建立一种有效、节约的河豚鱼肠道内容物总DNA提取方法,应用于聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)等分子生物学研究。方法:采取苯酚-氯仿-异戊醇抽提法,QIAamp DNA Stool Mini Kit,改良FastDNA SPIN Kit for Soil这3种DNA...目的:建立一种有效、节约的河豚鱼肠道内容物总DNA提取方法,应用于聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)等分子生物学研究。方法:采取苯酚-氯仿-异戊醇抽提法,QIAamp DNA Stool Mini Kit,改良FastDNA SPIN Kit for Soil这3种DNA提取方法,并结合PCR-DGGE技术分析河豚鱼肠道微生物多样性。结果:苯酚-氯仿-异戊醇抽提法与QIAamp DNA Stool Mini Kit提取的DNA量远高于改良FastDNA SPIN Kit forSoil,并且包含的微生物多样性较高。结论:苯酚-氯仿-异戊醇抽提法作为一种有效、节约的DNA提取方法,可应用于肠道微生物多样性研究。展开更多
Microbial community diversities in the drainage from three mines(Dexing Copper Mine,Qibaoshan Copper Mine and Yaogangxian Tungsten Mine,China)were analyzed using 16S rDNA PCR-RFLP approach.The efficiencies of chalcopy...Microbial community diversities in the drainage from three mines(Dexing Copper Mine,Qibaoshan Copper Mine and Yaogangxian Tungsten Mine,China)were analyzed using 16S rDNA PCR-RFLP approach.The efficiencies of chalcopyrite bioleaching were compared using enrichment of the three cultures.Phylogenetic analysis indicates that the dominant microorganisms are clustered with the Proteobacteria,the remaining is affiliated with Nitrospira,Acidobacteria and Actinobacteria.At the genus level,Acidithiobacillus is the dominant group in both YTW and QBS samples,while Spingomonas is dominant in YGX sample.Moreover,the principal component analysis(PCA)reveals that QBS and YTW have similar geochemical character and microbial communities.The results also show that pH value and tungsten concentration play a key role in microbial community distribution and relative abundance.The bioleaching efficiency of the enrichment cultures from YTW and QBS is similar.After 15 d,the bioleaching rates of low grade chalcopyrite(0.99%)are both up to 99.5% when using 10 g/L pulp density due to the similar microbial composition of YTW and QBS.Moreover,the leaching efficiencies of enrichment cultures containing multiple bioleaching microorganisms are higher than that of pure culture Acidithiobacillus ferrooxidans.展开更多
文摘目的:建立一种有效、节约的河豚鱼肠道内容物总DNA提取方法,应用于聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)等分子生物学研究。方法:采取苯酚-氯仿-异戊醇抽提法,QIAamp DNA Stool Mini Kit,改良FastDNA SPIN Kit for Soil这3种DNA提取方法,并结合PCR-DGGE技术分析河豚鱼肠道微生物多样性。结果:苯酚-氯仿-异戊醇抽提法与QIAamp DNA Stool Mini Kit提取的DNA量远高于改良FastDNA SPIN Kit forSoil,并且包含的微生物多样性较高。结论:苯酚-氯仿-异戊醇抽提法作为一种有效、节约的DNA提取方法,可应用于肠道微生物多样性研究。
基金Project(50621063) supported by the National Natural Science Foundation of ChinaProject (2004CB619204) supported by the National Basic Research and Development Program of China
文摘Microbial community diversities in the drainage from three mines(Dexing Copper Mine,Qibaoshan Copper Mine and Yaogangxian Tungsten Mine,China)were analyzed using 16S rDNA PCR-RFLP approach.The efficiencies of chalcopyrite bioleaching were compared using enrichment of the three cultures.Phylogenetic analysis indicates that the dominant microorganisms are clustered with the Proteobacteria,the remaining is affiliated with Nitrospira,Acidobacteria and Actinobacteria.At the genus level,Acidithiobacillus is the dominant group in both YTW and QBS samples,while Spingomonas is dominant in YGX sample.Moreover,the principal component analysis(PCA)reveals that QBS and YTW have similar geochemical character and microbial communities.The results also show that pH value and tungsten concentration play a key role in microbial community distribution and relative abundance.The bioleaching efficiency of the enrichment cultures from YTW and QBS is similar.After 15 d,the bioleaching rates of low grade chalcopyrite(0.99%)are both up to 99.5% when using 10 g/L pulp density due to the similar microbial composition of YTW and QBS.Moreover,the leaching efficiencies of enrichment cultures containing multiple bioleaching microorganisms are higher than that of pure culture Acidithiobacillus ferrooxidans.
