Identification and Characterization of a Gene stp17 Located on the Linear Plasmid pBSSB1 as an Enhanced Gene of Growth and Motility in Salmonella enterica Serovar Typhi.
Identification and Characterization of a Gene stp17 Located on the Linear Plasmid pBSSB1 as an Enhanced Gene of Growth and Motility in Salmonella enterica Serovar Typhi.
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位于线性质粒 pBSSB1 上的基因 stp17 的鉴定和表征，作为肠伤寒沙门氏菌生长和运动的增强基因
DOI：
10.3389/fcimb.2016.00110
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发表时间：
2016
影响因子：
5.7
通讯作者：
Huang X
中科院分区：
文献类型：
--
作者：
Zhang H;Zhu Y;Xie X;Wang M;Du H;Xu S;Zhang Y;Gong M;Ni B;Xu H;Huang X
线状表达载体pBSSB1介导了H：z66阳性伤寒沙门氏菌的鞭毛相变异。该基因命名为stp17(伤寒沙门氏菌质粒号17基因)，位于pBSSB1上，编码STP17蛋白。STP17在蛋白水平和功能上的表达模式尚不清楚。本研究对重组蛋白STP17His6进行了表达、纯化，并用于制备抗STP17的多克隆抗体。我们检测了stp17在伤寒沙门氏菌中的蛋白水平表达，并通过蛋白质印迹分析进一步研究了stp17在不同生长阶段的蛋白表达特征。分析了STP17对细菌生长和运动的影响。此外，对STP17的结构进行了预测，并通过定点突变鉴定了其活性部位。结果表明，STP17在伤寒沙门氏菌野生型菌株中稳定表达。当培养的OD600值达到1.2时，STP17在蛋白质水平的表达达到顶峰。与野生型相比，Δstp17株的生长速度和运动能力显著降低，而在stp17互补株中恢复了这一表型。此外，stp17过表达菌株的生长速度和活力均高于野生型菌株。STP17包括9个螺旋管段、6个支架管段和一些二级结构水平的线圈管段。I-tasser预测的STP17的顶端三维结构含有一个假定的ATPase结构域，GLY16、GLY19、LYS20、ASN133、LYS157和LYS158的氨基酸残基可能是STP17的活性位点残基。最后，STP17能够催化ATP合成ADP的反应，提示STP17可能是一种ATPase。据我们所知，这是首次描述STP17蛋白在伤寒沙门氏菌中的表达特征，表明STP17促进细菌的生长和运动，这可能与其潜在的ATPase活性有关。
The linear plasmid pBSSB1 mediates the flagellar phase variation in H:z66 positive Salmonella enterica serovar Typhi (S. Typhi). The gene named stp17 (S. Typhi plasmid number 17 gene) is located on pBSSB1 and encodes the protein STP17. The expression pattern at the protein-level and function of STP17 remains unknown. In this study, the recombinant protein STP17His6 was expressed, purified and used to prepare the polyclonal anti-STP17 antibody. We detected protein-level expression of stp17 in S. Typhi and further investigated the protein expression characteristics of stp17 in different growth phases by western blot analysis. The effects of STP17 on bacterial growth and motility were analyzed. In addition, the structure of STP17 was predicted and the active site of STP17 was identified by site-directed mutagenesis. The results showed that STP17 was expressed stably in the wild type strain of S. Typhi. STP17 expression at the protein level peaks when cultures reach an OD600 value of 1.2. The growth rate and motility of the Δstp17 strain were significantly decreased compared with the wild type strain (P < 0.05) and this phenotype was restored in the stp17 complementary strain. Moreover, the growth rate and motility of the stp17 over-expression strain was greater than the wild type strain. STP17 contains nine Helix segments, six Stand segments and some Coil segments in the secondary structural level. The top-ranked 3-D structure of STP17 predicted by I-TASSER contains a putative ATPase domain and the amino acid residues of GLY16, GLY19, LYS20, ASN133, LYS157, and LYS158 may be the active site residues of STP17. Finally, STP17 was able to catalyze the ATP to ADP reaction, suggesting that STP17 may be an ATPase. To our knowledge, this is the first report describing the protein expression characteristics of STP17 in S. Typhi, showing that STP17 promotes bacterial growth and motility, which may be associated with its potential ATPase activity.
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影响因子：
6.7
作者：
Baker, Stephen;Hardy, Jonathan;Sanderson, Kenneth E;Quail, Michael;Goodhead, Ian;Kingsley, Robert A;Parkhill, Julian;Stocker, Bruce;Dougan, Gordon
通讯作者：
Dougan, Gordon
影响因子：
64.8
作者：
PLASTERK, RHA;SIMON, MI;BARBOUR, AG
通讯作者：
BARBOUR, AG
影响因子：
4.6
作者：
Bai F;Morimoto YV;Yoshimura SD;Hara N;Kami-Ike N;Namba K;Minamino T
通讯作者：
Minamino T
DOI：
10.1590/s1517-83822010000300025
发表时间：
2010-07
期刊：
Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]
影响因子：
--
作者：
Xu S;Zou X;Sheng X;Zhang H;Mao L;Du H;Xu H;Huang X
通讯作者：
Huang X
影响因子：
2.6
作者：
El-Mansi, M;Anderson, KJ;Platt, DJ
通讯作者：
Platt, DJ