This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Shen, J.
Right arrow Articles by Zhao, L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Shen, J.
Right arrow Articles by Zhao, L.
Agricola
Right arrow Articles by Shen, J.
Right arrow Articles by Zhao, L.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, August 2006, p. 5232-5238, Vol. 72, No. 8
0099-2240/06/$08.00+0     doi:10.1128/AEM.00151-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Molecular Profiling of the Clostridium leptum Subgroup in Human Fecal Microflora by PCR-Denaturing Gradient Gel Electrophoresis and Clone Library Analysis

Jian Shen,1 Baorang Zhang,2 Guifang Wei,1 Xiaoyan Pang,1 Hua Wei,1 Min Li,1 Yan Zhang,1 Wei Jia,3 and Liping Zhao1*

Laboratory of Molecular Microbial Ecology and Ecogenomics, College of Life Science and Biotechnology, Shanghai Jiaotong University, Shanghai 200240, China,1 Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Shanxi University, Shanxi 030006, China,2 School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China3

Received 20 January 2006/ Accepted 24 May 2006

A group-specific PCR-based denaturing gradient gel electrophoresis (DGGE) method was developed and combined with group-specific clone library analysis to investigate the diversity of the Clostridium leptum subgroup in human feces. PCR products (length, 239 bp) were amplified using C. leptum cluster-specific primers and were well separated by DGGE. The DGGE patterns of fecal amplicons from 11 human individuals revealed host-specific profiles; the patterns for fecal samples collected from a child for 3 years demonstrated the structural succession of the population in the first 2 years and its stability in the third year. A clone library was constructed with 100 clones consisting of 1,143-bp inserts of 16S rRNA gene fragments that were amplified from one adult fecal DNA with one forward universal bacterial primer and one reverse group-specific primer. Eighty-six of the clones produced the 239-bp C. leptum cluster-specific amplicons, and the remaining 14 clones did not produce these amplicons but still phylogenetically belong to the subgroup. Sixty-four percent of the clones were related to Faecalibacterium prausnitzii (similarity, 97 to 99%), 6% were related to Subdoligranulum variabile (similarity, ~99%), 2% were related to butyrate-producing bacterium A2-207 (similarity, 99%), and 28% were not identified at the species level. The identities of most bands in the DGGE profiles for the same adult were determined by comigration analysis with the 86 clones that harbored the 239-bp group-specific fragments. Our results suggest that DGGE combined with clone library analysis is an effective technique for monitoring and analyzing the composition of this important population in the human gut flora.

* Corresponding author. Mailing address: Laboratory of Molecular Microbial Ecology and Ecogenomics, College of Life Science and Biotechnology, Shanghai Jiaotong University, Shanghai 200240, China. Phone: 86 21 34204877 or 86 21 34204965. Fax: 86 21 34204877. E-mail: lpzhao{at}sjtu.edu.cn.

Applied and Environmental Microbiology, August 2006, p. 5232-5238, Vol. 72, No. 8
0099-2240/06/$08.00+0     doi:10.1128/AEM.00151-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Julien, M.-C., Dion, P., Lafreniere, C., Antoun, H., Drouin, P. (2008). Sources of Clostridia in Raw Milk on Farms. Appl. Environ. Microbiol. 74: 6348-6357 [Abstract] [Full Text]  
  • Li, M., Wang, B., Zhang, M., Rantalainen, M., Wang, S., Zhou, H., Zhang, Y., Shen, J., Pang, X., Zhang, M., Wei, H., Chen, Y., Lu, H., Zuo, J., Su, M., Qiu, Y., Jia, W., Xiao, C., Smith, L. M., Yang, S., Holmes, E., Tang, H., Zhao, G., Nicholson, J. K., Li, L., Zhao, L. (2008). Symbiotic gut microbes modulate human metabolic phenotypes. Proc. Natl. Acad. Sci. USA 105: 2117-2122 [Abstract] [Full Text]  
  • Kuramitsu, H. K., He, X., Lux, R., Anderson, M. H., Shi, W. (2007). Interspecies Interactions within Oral Microbial Communities. Microbiol. Mol. Biol. Rev. 71: 653-670 [Abstract] [Full Text]  
  • Zhang, M., Liu, B., Zhang, Y., Wei, H., Lei, Y., Zhao, L. (2007). Structural Shifts of Mucosa-Associated Lactobacilli and Clostridium leptum Subgroup in Patients with Ulcerative Colitis. J. Clin. Microbiol. 45: 496-500 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»