Screening of genes regulated by cold shock in Shewanella piezotolerans WP3 and time course expression of cold-regulated genes
Screening of genes regulated by cold shock in Shewanella piezotolerans WP3 and time course expression of cold-regulated genes
复制标题
压电希瓦氏菌WP3冷激调控基因的筛选及冷调控基因的时程表达
DOI：
10.1007/s00203-007-0347-1
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发表时间：
2008-06-01
影响因子：
2.8
通讯作者：
Wang, Fengping
中科院分区：
文献类型：
--
作者：
Li, Shengkang;Xiao, Xiang;Wang, Fengping
利用RNA任意引物PCR技术分析了深海细菌Shewanella piezotolerans WP 3在冷激条件下的差异基因转录。使用90个引物组扫描来自冷休克培养物及其对照（在其最佳生长温度下培养）的两个不同RNA库。鉴定并克隆了94个推定的差异表达片段。经反向北方斑点杂交证实，94个片段中有6个确实是冷休克差异转录片段，并进行了测序。序列比对分析表明，这6个差异转录基因分别是小带封闭毒素、伴侣GroEL、外排转运蛋白、Sua 5/YciO/YrdC/YwlC家族蛋白、甜菜碱醛脱氢酶和DEAD盒RNA解旋酶的推定基因。通过实时定量PCR对冷休克后0 ~ 90 min这6个基因的时程表达谱进行定量。最高诱导基因--RNA解旋酶基因的缺失突变，无论是在冷激条件下还是在持续低温条件下，对菌株的生长均无显著影响。这表明，一个或多个额外的DEAD盒RNA解旋酶基因补偿突变基因的功能丧失。
The differential gene transcription of a deep-sea bacterium Shewanella piezotolerans WP3 in response to cold shock was analyzed by RNA arbitrarily primed PCR. Ninety primer sets were used to scan two different RNA pools derived from the culture of cold shock and its control (culture at its optimal grown temperature). Ninety-four putative differentially expressed fragments were identified and cloned. Six out of the 94 fragments were confirmed to be truly differentially transcribed in terms of cold shock by reverse Northern dot blot and then sequenced. Sequence blast analysis showed that the six differentially transcribed genes are putative genes for zonular occludens toxin, chaperon GroEL, efflux transporter, Sua5/YciO/YrdC/YwlC family protein, betaine-aldehyde dehydrogenase, and DEAD box RNA helicase, respectively. The time course expression profiles of these six genes from 0 to 90 min upon cold shock were quantified by real-time PCR. Deletion mutation of the highest induced gene--RNA helicase gene, had no significant impact on the growth of the strain no matter upon cold shock or under permanent low temperature. It is suggested that one or more additional DEAD box RNA helicase genes compensate for the loss of the function of the mutated gene.