Identification of a large motility operon in Borrelia burgdorferi by semi-random PCR chromosome walking

Gene
Y Ge, Nyles W Charon

Abstract

Motility has been implicated in the invasive process of Borrelia burgdorferi (Bb), the etiologic agent of Lyme disease. To identify Bb motility related genes, we used a method termed 'semi-random PCR chromosome walking' (SRPCW) to walk through a large motility gene cluster. The major advantage of this approach over other PCR walking methods is that it employs a secondary PCR amplification of cloned fragments which can be readily sequenced and analyzed. Starting with a primer specific to flgE, we identified and sequenced 14 open reading frames (ORFs) spanning 11 kb downstream of the flgE gene. The genes identified include flbD, motA, motB, fliL, fliM, fliN, fliZ, fliP, fliQ, fliR, flhB, flhA, flhF and flbE. Twelve of the deduced proteins shared extensive homology with flagellar proteins from other bacteria. The gene products and order of genes within this cluster are most similar to those of Treponema pallidum (Tp) and Bacillus subtilis (Bs). One of the unique genes identified, flbD, demonstrated homology to an ORF from the same operon of Tp. Another ORF, flbE, showed similarity to genes from both Tp and Bs. RT-PCR and primer extension analysis revealed that this gene cluster is transcribed as a single unit indicating that it is...Continue Reading

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Citations

Aug 29, 2009·Applied Microbiology and Biotechnology·Yuki Tonooka, Masahiro Fujishima
Feb 3, 2005·Nature Reviews. Microbiology·Patricia A RosaPhilip E Stewart
Aug 14, 2013·Proceedings of the National Academy of Sciences of the United States of America·Xiaowei ZhaoJun Liu
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Nov 22, 2016·Nature Microbiology·Kai ZhangChunhao Li
Oct 22, 2020·Journal, Genetic Engineering & Biotechnology·Seyedeh-Samira AshrafmansouriMarie Seidi

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