Inactivation of the Flagellin Gene flaA in Magnetospirillum gryphiswaldense Results in Nonmagnetotactic Mutants Lacking Flagellar Filaments
Inactivation of the Flagellin Gene flaA in Magnetospirillum gryphiswaldense Results in Nonmagnetotactic Mutants Lacking Flagellar Filaments
复制标题
磁螺菌 gryphiswaldense 中鞭毛蛋白基因 flaA 的失活导致缺乏鞭毛丝的非趋磁突变体
DOI：
10.1128/aem.70.6.3624-3631.2004
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发表时间：
2004
影响因子：
4.4
通讯作者：
D. Schüler
中科院分区：
文献类型：
--
作者：
D. Schultheiss;M. Kube;D. Schüler
趋磁细菌合成磁小体，使其沿磁力线定向和迁移。在分子水平上分析趋磁性和磁小体生物矿化一直受到遗传方法不可用性的阻碍，即缺乏引入定向基因特异性突变的手段。本文报道了一种用同源重组法对gryphiswaldense磁螺旋藻进行基因敲除诱变的方法。在M. gryphiswaldense中发现了多个鞭毛蛋白基因，这些基因在基因组中未连接。鞭毛蛋白基因flaA的靶向破坏被证明可以消除鞭毛的形成、运动性和趋磁性。本文描述的技术将使充分利用即将到来的gryphiswaldense和其他趋磁细菌的基因组序列成为可能。
ABSTRACT Magnetotactic bacteria synthesize magnetosomes, which cause them to orient and migrate along magnetic field lines. The analysis of magnetotaxis and magnetosome biomineralization at the molecular level has been hindered by the unavailability of genetic methods, namely the lack of a means to introduce directed gene-specific mutations. Here we report a method for knockout mutagenesis by homologous recombination in Magnetospirillum gryphiswaldense. Multiple flagellin genes, which are unlinked in the genome, were identified in M. gryphiswaldense. The targeted disruption of the flagellin gene flaA was shown to eliminate flagella formation, motility, and magnetotaxis. The techniques described in this paper will make it possible to take full advantage of the forthcoming genome sequences of M. gryphiswaldense and other magnetotactic bacteria.