Approach to analyze the diversity of myxobacteria in soil by semi-nested PCR-denaturing gradient gel electrophoresis (DGGE) based on taxon-specific gene.
Approach to analyze the diversity of myxobacteria in soil by semi-nested PCR-denaturing gradient gel electrophoresis (DGGE) based on taxon-specific gene.
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DOI：
10.1371/journal.pone.0108877
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Zhu H
中科院分区：
文献类型：
--
作者：
Li B;Yao Q;Zhu H
不溶性大分子的基因多样性降解了粘细菌，为发现新的细菌资源和发现新的生态功能提供了机会。在这项研究中，我们建立了一种半套式-聚合酶链式反应-变性梯度凝胶电泳法(DGGE)来检测土壤中粘细菌的存在和基因多样性。用粘细菌特异性引物进行两轮聚合酶链式反应，获得了适合于DGGE的滑动运动关键基因mglA的194个核苷酸片段。在DGGE图谱中观察到大量的条带，表明土壤中存在着多样性的粘细菌。测序和BLAST分析表明，大部分条带属于粘细菌类群，28条条带中只有3条属于另一类群，即大理链球菌。结果表明，用粘细菌特异的DGGE方法可以区分mglA基因序列组成不同的粘细菌。总之，开发的半套式-PCR-DGGE策略是研究粘细菌多样性的有用工具。
The genotypic diversity of insoluble macromolecules degraded myxobacteria, provided an opportunity to discover new bacterial resources and find new ecological functions. In this study, we developed a semi-nested-PCR-denaturing gradient gel electrophoresis (DGGE) strategy to determine the presence and genotypic diversity of myxobacteria in soil. After two rounds of PCR with myxobacteria-specific primers, an 194 bp fragment of mglA, a key gene involved in gliding motility, suitable for DGGE was obtained. A large number of bands were observed in DGGE patterns, indicating diverse myxobacteria inhabiting in soils. Furthermore, sequencing and BLAST revealed that most of the bands belonged to the myxobacteria-group, and only three of the twenty-eight bands belonged to other group, i.e., Deinococcus maricopensis. The results verified that myxobacterial strains with discrepant sequence compositions of gene mglA could be discriminated by DGGE with myxobacteria-specific primers. Collectively, the developed semi-nested-PCR-DGGE strategy is a useful tool for studying the diversity of myxobacteria.
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DOI：
10.1073/pnas.82.24.8767
发表时间：
1985-12-01
影响因子：
11.1
作者：
BLACKHART, BD;ZUSMAN, DR
通讯作者：
ZUSMAN, DR
影响因子：
4.4
作者：
Dar, SA;Kuenen, JG;Muyzer, G
通讯作者：
Muyzer, G
DOI：
10.1023/a:1000669317571
发表时间：
1998-01-01
影响因子：
2.6
作者：
Muyzer, G;Smalla, K
通讯作者：
Smalla, K
影响因子：
4.4
作者：
Salles, JF;De Souza, FA;van Elsas, JD
通讯作者：
van Elsas, JD
影响因子：
14.9
作者：
MURRAY, MG;THOMPSON, WF
通讯作者：
THOMPSON, WF