In order to reduce deleterious effect on environment,human health and facilities caused by original sulfides, more attention should be paid to biodesulfurization studying for fossil fuels. In this work, eight isolates...In order to reduce deleterious effect on environment,human health and facilities caused by original sulfides, more attention should be paid to biodesulfurization studying for fossil fuels. In this work, eight isolates were characterized by several DNA-based methods such as BOX element polymerase chain reaction( BOX-PCR), enterobacterial repetitive intergenic consensus( ERIC)-PCR and random amplification of polymorphic DNA( RAPD)-PCR. The desulfurization performance was determined by micro-coulometric method,Gibb's assay and barium sulfate test. It was found out that ERIC-PCR displays a much higher inter-strain heterogeneity compared with using BOX. The length of the primer didnot play the most important role in bacterial classification. The combination of the analysis of repetitive-sequence-based polymerase chain reaction ngerprinting and 16 S r DNA was able to provide more effective way in the separation and identification of bacteria.According to the analysis of 16 S r DNA,the more efficient desulfurization strain should belong to Klebsiella variicola.展开更多
目的:肠道微生物的研究日益成为热点,如何获取高质量、较完整的肠道菌群基因组DNA是肠道微生物研究中的关键。本文通过对酚/氯仿法提取总DNA过程进行考察和优化,建立一种简便酚/氯仿抽提法。方法:考察和优化酚/氯仿法提取总DNA的过程,...目的:肠道微生物的研究日益成为热点,如何获取高质量、较完整的肠道菌群基因组DNA是肠道微生物研究中的关键。本文通过对酚/氯仿法提取总DNA过程进行考察和优化,建立一种简便酚/氯仿抽提法。方法:考察和优化酚/氯仿法提取总DNA的过程,并根据DNA产量、纯度以及ERIC-PCR及16S rNDA-RFLP所反映的微生物群落结构特性的指标,并与QIAamp?DNA Stool Mini Kit提取的进行比较,评价了所建立的快速提取方法。结果:用简便酚/氯仿法得到基本完整的基因组DNA,ERIC-PCR和16S rDNA-RFLP结果与QIAamp?DNA Stool Mini Kit法基本相同。结论:该方法快速并成本低,适合肠道微生物研究中总DNA提取,尤其适合处理大批量的样品。展开更多
随着现代分子生物学技术的快速发展,动物胃肠道微生物领域的研究技术也进入了一个新的阶段。文章从传统及现代分子生物学两方面介绍了几种常用的研究动物胃肠道微生物技术及其应用进展,主要包括:传统微生物培养技术、16S r DNA序列分析...随着现代分子生物学技术的快速发展,动物胃肠道微生物领域的研究技术也进入了一个新的阶段。文章从传统及现代分子生物学两方面介绍了几种常用的研究动物胃肠道微生物技术及其应用进展,主要包括:传统微生物培养技术、16S r DNA序列分析技术、16S rRNA序列分析技术[变性梯度凝胶电泳(DGGE)、温度梯度凝胶电泳(TGGE)、单链构象多肽性(PCR-SSCP)、末段限制性长度多态性(T-RFLP)]、ERIC-PCR技术、RAPD技术等,并对动物肠道微生物研究技术的发展进行了展望。展开更多
基金National Natural Science Foundations of China(Nos.41302079,21176145)Project of Shandong Province Higher Educational Science and Technology Program,China(No.J13LD54)
文摘In order to reduce deleterious effect on environment,human health and facilities caused by original sulfides, more attention should be paid to biodesulfurization studying for fossil fuels. In this work, eight isolates were characterized by several DNA-based methods such as BOX element polymerase chain reaction( BOX-PCR), enterobacterial repetitive intergenic consensus( ERIC)-PCR and random amplification of polymorphic DNA( RAPD)-PCR. The desulfurization performance was determined by micro-coulometric method,Gibb's assay and barium sulfate test. It was found out that ERIC-PCR displays a much higher inter-strain heterogeneity compared with using BOX. The length of the primer didnot play the most important role in bacterial classification. The combination of the analysis of repetitive-sequence-based polymerase chain reaction ngerprinting and 16 S r DNA was able to provide more effective way in the separation and identification of bacteria.According to the analysis of 16 S r DNA,the more efficient desulfurization strain should belong to Klebsiella variicola.
文摘目的:肠道微生物的研究日益成为热点,如何获取高质量、较完整的肠道菌群基因组DNA是肠道微生物研究中的关键。本文通过对酚/氯仿法提取总DNA过程进行考察和优化,建立一种简便酚/氯仿抽提法。方法:考察和优化酚/氯仿法提取总DNA的过程,并根据DNA产量、纯度以及ERIC-PCR及16S rNDA-RFLP所反映的微生物群落结构特性的指标,并与QIAamp?DNA Stool Mini Kit提取的进行比较,评价了所建立的快速提取方法。结果:用简便酚/氯仿法得到基本完整的基因组DNA,ERIC-PCR和16S rDNA-RFLP结果与QIAamp?DNA Stool Mini Kit法基本相同。结论:该方法快速并成本低,适合肠道微生物研究中总DNA提取,尤其适合处理大批量的样品。
文摘随着现代分子生物学技术的快速发展,动物胃肠道微生物领域的研究技术也进入了一个新的阶段。文章从传统及现代分子生物学两方面介绍了几种常用的研究动物胃肠道微生物技术及其应用进展,主要包括:传统微生物培养技术、16S r DNA序列分析技术、16S rRNA序列分析技术[变性梯度凝胶电泳(DGGE)、温度梯度凝胶电泳(TGGE)、单链构象多肽性(PCR-SSCP)、末段限制性长度多态性(T-RFLP)]、ERIC-PCR技术、RAPD技术等,并对动物肠道微生物研究技术的发展进行了展望。
