Exploration of chlamydial type III secretion system reconstitution in Escherichia coli.
Exploration of chlamydial type III secretion system reconstitution in Escherichia coli.
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DOI：
10.1371/journal.pone.0050833
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Fan H
中科院分区：
文献类型：
--
作者：
Bao X;Beatty WL;Fan H
III型分泌系统是许多病原体的毒力因子，并且被认为在衣原体（Chlamydia）的发育周期和致病性中起多个作用，衣原体是重要的人类病原体。然而，由于衣原体的专性细胞内寄生性质和缺乏方便的遗传学方法的生物体，非常有限的方法可用于研究衣原体III型分泌系统。在这项研究中，我们探讨了重组衣原体III型分泌大肠杆菌。我们成功地将衣原体III型分泌系统的所有6个基因组DNA簇克隆到三个细菌质粒中。发现6个簇中的5个簇在用三种质粒转化的大肠杆菌中从它们自己的启动子指导mRNA合成。簇5未能使用其自身的启动子表达mRNA。然而，簇5与簇6的融合导致簇5mRNA的表达。虽然只有两个III型分泌系统蛋白被检测到转化E。大肠杆菌中，由于有限的抗体可用性，III型分泌系统样结构中检测到的转化E.杆菌我们已经成功地生成了E。表达衣原体III型分泌系统的所有基因的大肠杆菌。这为重组衣原体III型分泌系统的最佳表达和组装奠定了基础，这对于衣原体III型分泌系统的表征和研究其在衣原体致病性中的作用可能非常有用。
Type III secretion system is a virulent factor for many pathogens, and is thought to play multiple roles in the development cycle and pathogenesis of chlamydia, an important human pathogen. However, due to the obligate intracellular parasitical nature of chlamydiae and a lack of convenient genetic methodology for the organisms, very limited approaches are available to study the chlamydial type III secretion system. In this study, we explored the reconstitution of a chlamydial type III secretion in Escherichia coli. We successfully cloned all 6 genomic DNA clusters of the chlamydial type III secretion system into three bacterial plasmids. 5 of the 6 clusters were found to direct mRNA synthesis from their own promoters in Escherichia coli transformed with the three plasmids. Cluster 5 failed to express mRNA using its own promoters. However, fusion of cluster 5 to cluster 6 resulted in the expression of cluster 5 mRNA. Although only two of the type III secretion system proteins were detected transformed E. coli due to limited antibody availability, type III secretion system-like structures were detected in ultrathin sections in a small proportion of transformed E. coli. We have successfully generated E. coli expressing all genes of the chlamydial type III secretion system. This serves as a foundation for optimal expression and assembly of the recombinant chlamydial type III secretion system, which may be extremely useful for the characterization of the chlamydial type III secretion system and for studying its role in chlamydial pathogenicity.
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影响因子：
3.6
作者：
Fields, KA;Hackstadt, T
通讯作者：
Hackstadt, T
DOI：
10.1073/pnas.0603044103
发表时间：
2006-10-17
影响因子：
11.1
作者：
Jewett, Travis J.;Fischer, Elizabeth R.;Hackstadt, Ted
通讯作者：
Hackstadt, Ted
无细胞沙眼衣原体原体诱导 III 型分泌。
影响因子：
3.8
作者：
Jamison, Wendy P.;Hackstadt, Ted
通讯作者：
Hackstadt, Ted
DOI：
10.1073/pnas.89.13.5976
发表时间：
1992-07-01
影响因子：
11.1
作者：
GINOCCHIO, C;PACE, J;GALAN, JE
通讯作者：
GALAN, JE
DOI：
10.1099/mic.0.049668-0
发表时间：
2011-09
期刊：
Microbiology (Reading, England)
影响因子：
--
作者：
Bao X;Pachikara ND;Oey CB;Balakrishnan A;Westblade LF;Tan M;Chase T;Nickels BE;Fan H
通讯作者：
Fan H