Gas vesicle genes identified in Bacillus megaterium and functional expression in Escherichia coli
Gas vesicle genes identified in Bacillus megaterium and functional expression in Escherichia coli
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DOI：
10.1128/jb.180.9.2450-2458.1998
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发表时间：
1998-05-01
影响因子：
3.2
通讯作者：
Cannon, MC
中科院分区：
文献类型：
--
作者：
Li, N;Cannon, MC
气泡是在蓝细菌和嗜盐古细菌中发现的细胞内的、蛋白质包被的和中空的细胞器。它们通过扩散可渗透周围气体并提供浮力，使细胞能够在液体中向上移动以获得氧气和/或光。在盐细菌中，气泡的产生是由14个基因（4个已知功能）组成的9 kb簇编码的。在蓝藻中，涉及的基因数量尚未确定。我们现在报告的克隆和序列分析的8,142 bp的15个推定的气体囊泡基因（gvp）从巨大芽孢杆菌VT 1660和它们的功能表达在大肠杆菌。证据包括通过序列分析与已知的气泡基因同源，浮力表型关闭。携带该grp基因簇的大肠杆菌菌株，相差显微镜中压力敏感的可触小体的存在，相差显微镜中的结构细节，直接干涉-相差显微镜中的结构细节，以及透射电子显微镜显示的形状和大小。在B。在巨大芽孢杆菌中，gvp区携带一个由15个假定基因组成的簇，它们是开放阅读框1和gvpA、-P、-Q、-B、-R、-N、-F、-G、-L、-S、-K、-J、-T和-U，其中最后的II基因在5.7-kb基因簇中，是E中气泡合成和功能所需的最大基因。杆菌据我们所知，这是非水生细菌中功能性气泡基因簇的第一个例子，也是导致功能性细胞器合成的基因种间转移的第一个例子。
Gas vesicles are intracellular, protein-coated, and hollow organelles found in cyanobacteria and halophilic archaea. They are permeable to ambient gases by diffusion and provide buoyancy, enabling cells to move upwards in liquid to access oxygen and/or light. In halobacteria, gas vesicle production is encoded in a 9-kb cluster of 14 genes (4 of known function). In cyanobacteria, the number of genes involved has not been determined. We now report the cloning and sequence analysis of an 8,142-bp cluster of 15 putative gas vesicle genes (gvp) from Bacillus megaterium VT1660 and their functional expression in Escherichia coli. Evidence includes homologies by sequence analysis to known gas vesicle genes, the buoyancy phenotype off. coli strains that carry this grp gene cluster, the presence of pressure-sensitive, refractile bodies in phase contrast microscopy, structural details in phase-constrast microscopy, structural details in direct interference-contrast microscopy, and shape and size revealed by transmission electron microscopy. In B. megaterium, the gvp region carries a cluster of 15 putative genes arranged in one orientation; they are open reading frame 1 and gvpA, -P, -Q, -B, -R, -N, -F, -G, -L, -S, -K, -J, -T, and -U, of which the last II genes, in a 5.7-kb gene cluster, are the maximum required for gas vesicle synthesis and function in E. coli. To our knowledge, this Is the first example of a functional gas vesicle gene cluster in nonaquatic bacteria and the first example of the interspecies transfer of genes resulting in the synthesis of a functional organelle.