Identification of genes regulated by changing salinity in the deep-sea bacterium Shewanella sp WP3 using RNA arbitrarily primed PCR
Identification of genes regulated by changing salinity in the deep-sea bacterium Shewanella sp WP3 using RNA arbitrarily primed PCR
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DOI：
10.1007/s00792-005-0476-x
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发表时间：
2006-04-01
期刊：
影响因子：
2.9
通讯作者：
Wang, FP
中科院分区：
文献类型：
--
作者：
Li, SK;Xiao, X;Wang, FP
采用随机引物PCR技术（RAP-PCR）分析了深海细菌希瓦氏菌（Shewanella sp.）WP 3在盐度变化中的基因转录差异。使用90个引物组来扫描来自1%和7%NaCl浓度的培养物的两个不同的RNA池。对43个推定的差异表达片段进行鉴定、克隆和测序。在43个片段中，有6个片段被证实在盐度变化方面是真正差异转录的。这6个基因片段的氨基酸序列与Shewanella oneidensis MR-1的核糖体蛋白L24、ATP结合蛋白和伴侣蛋白HscA的同源性最高（66- 96（Y 6、Y 9和Y29）;铜绿假单胞菌的异柠檬酸裂解酶（Y15）;希瓦氏菌SIB 1的肽基脯氨酰顺反异构酶（Y21）、紫色希瓦氏菌谷氨酰胺合成酶（Y25）。4个基因（Y 6、Y15、Y21和Y25）在7%NaCl中表达上调，而另外2个基因（Y 9和Y29）在1%NaCl中表达量更高。这些数据表明，涉及控制蛋白质合成，蛋白质折叠和/或贩运，谷氨酸浓度，脂肪酸代谢和物质转运的策略被用于希瓦氏菌的盐适应。进一步检查了响应瞬时应力冲击的六个基因的表达模式，包括盐冲击（3%NaCl转变为12%）、冷冲击（15 ℃转变为0 ℃）和高静水压力冲击（0.1 MPa转变为50 MPa）。编码推定的HscA伴侣蛋白的Y29被指示参与所有测试的应力的适应。
The differential gene transcription of a deep-sea bacterium Shewanella sp. WP3 in response to changing salinity was analyzed by RNA fingerprinting using arbitrarily primed PCR (RAP-PCR). Ninety primer sets were used to scan two different RNA pools derived from cultures of 1% and 7% NaCl concentrations. Forty-three putative differential-expressed fragments were identified, cloned, and sequenced. Six out of the 43 fragments were confirmed to be truly differentially transcribed in terms of changing salinity. The deduced amino acid sequences of the six gene fragments showed highest identities (66-96%) with ribosomal protein L24, ATP binding protein, and chaperon protein HscA of Shewanella oneidensis MR-1 (Y6, Y9, and Y29); isocitrate lyase of Pseudomonas aeruginosa (Y15); peptidylprolyl cis-trans isomerase of Shewanella sp. SIB1 (Y21), glutamine synthetase of Shewanella violacea (Y25), respectively. Four genes (Y6, Y15, Y21, and Y25) were up regulated in 7% NaCl, while the other two (Y9 and Y29) contained more abundant transcripts in 1% NaCl. The data suggested that strategies involved in controlling protein synthesis, protein folding and/or trafficking, glutamate concentration, fatty acid metabolism, and substance transporting were used for salt adaptation in Shewanella sp. WP3. The expression patterns of the six genes in response to transient stress shocks including salt shock (3% NaCl shift to 12%), cold shock (15 degrees C shift to 0 degrees C), and high-hydrostatic pressure shock (0.1 MPa shift to 50 MPa) were further examined. Y29 encoding the putative HscA chaperon protein was indicated to be involved in adaptation of all the stresses tested.