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

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, December 2005, p. 7995-8001, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.7995-8001.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Molecular Analysis of the O-Antigen Gene Cluster of Escherichia coli O86:B7 and Characterization of the Chain Length Determinant Gene (wzz)

Hongjie Guo,¹ Wen Yi,¹ Jun Shao,¹ Yuquan Lu,¹ Wenpeng Zhang,¹ Jing Song,² and Peng George Wang^1,2*

Department of Biochemistry and Chemistry, The Ohio State University, Columbus, Ohio 43210,¹ The State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Jinan, Shandong 250100, People's Republic of China²

Received 1 July 2005/ Accepted 11 August 2005

Escherichia coli O86:B7 has long been used as a model bacterial strain to study the generation of natural blood group antibody in humans, and it has been shown to possess high human blood B activity. The O-antigen structure of O86:B7 was solved recently in our laboratory. Comparison with the published structure of O86:H2 showed that both O86 subtypes shared the same O unit, yet each of the O antigens is polymerized from a different terminal sugar in a different glycosidic linkage. To determine the genetic basis for the O-antigen differences between the two O86 strains, we report the complete sequence of O86:B7 O-antigen gene cluster between galF and hisI, each gene was identified based on homology to other genes in the GenBank databases. Comparison of the two O86 O-antigen gene clusters revealed that the encoding regions between galF and gnd are identical, including wzy genes. However, deletion of the two wzy genes revealed that wzy in O86:B7 is responsible for the polymerization of the O antigen, while the deletion of wzy in O86:H2 has no effect on O-antigen biosynthesis. Therefore, we proposed that there must be another functional wzy gene outside the O86:H2 O-antigen gene cluster. Wzz proteins determine the degree of polymerization of the O antigen. When separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the lipopolysaccharide (LPS) of O86:B7 exhibited a modal distribution of LPS bands with relatively short O units attached to lipid A-core, which differs from the LPS pattern of O86:H2. We proved that the wzz genes are responsible for the different LPS patterns found in the two O86 subtypes, and we also showed that the very short type of LPS is responsible for the serum sensitivity of the O86:B7 strain.

---

* Corresponding author. Mailing address: Department of Biochemistry and Chemistry, The Ohio State University, Columbus, OH 43210. Phone: (614) 292-9884. Fax: (614) 292-3106. E-mail: wang.892{at}osu.edu.

---

Applied and Environmental Microbiology, December 2005, p. 7995-8001, Vol. 71, No. 12
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.12.7995-8001.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

This article has been cited by other articles:

• Yi, W., Liu, X., Li, Y., Li, J., Xia, C., Zhou, G., Zhang, W., Zhao, W., Chen, X., Wang, P. G. (2009). Remodeling bacterial polysaccharides by metabolic pathway engineering. Proc. Natl. Acad. Sci. USA 106: 4207-4212 [Abstract] [Full Text]  
• Iguchi, A., Ooka, T., Ogura, Y., Asadulghani, , Nakayama, K., Frankel, G., Hayashi, T. (2008). Genomic comparison of the O-antigen biosynthesis gene clusters of Escherichia coli O55 strains belonging to three distinct lineages. Microbiology 154: 559-570 [Abstract] [Full Text]