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        Screening and Identification of ClpE Interaction Proteins in Streptococcus pneumoniae by a Bacterial Two-Hybrid System and Co-immunoprecipitation

        WenJuan Yan,YingYing Cai,Qun Zhang,YuSi Liu,WenChun Xu,YiBing Yin,YuJuan He,Hong Wang,XueMei Zhang 한국미생물학회 2013 The journal of microbiology Vol.51 No.4

        Hsp100/Clp proteins have crucial functions in the protein quality control, stress tolerance, and virulence of many pathogenic bacteria. ClpE is an important virulence factor involved in adherence and invasion in Streptococcus pneumoniae. To explore the underlying mechanism, we screened ClpE interaction proteins using a bacterial two-hybrid system and co-immunoprecipitation. We used ClpE as bait and constructed the pBT-ClpE bait plasmid for two-hybrid screening. Then, we constructed ClpE::GFP fusion for co-immunoprecipitation screening using anti-GFP monoclonal antibody. We obtained eight potential ClpE interaction proteins,including carbamoyl-phosphate synthase, pyruvate oxidase (SpxB), phosphoenolpyruvate-protein phosphotransferase,aminopeptidase N (pepN), L-lactate dehydrogenase, ribosomal protein S4, sensor histidine kinase (SPD_2019), and FtsW (a cell division protein). FtsW, SpxB, pepN, and SPD_2019 were confirmed to interact with ClpE using Bacterial Two-hybrid or Co-immunoprecipitation. Morphologic observations found that ΔclpE strain existed in abnormal division. β-Galactosidase activity assay suggested that ClpE contributed to the degradation of FtsW. Furthermore, FtsW could be induced by heat shock. The results suggested that ClpE might affect cell division by regulating the level of FtsW. These data may provide new insights in studying the role of ClpE in S. pneumoniae.

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        Fluorescent Nitrogen-Doped Carbon Dots via Single-Step Synthesis Applied as Fluorescent Probe for the Detection of Fe3+ Ions and Anti-Counterfeiting Inks

        Chonghui Fan,Kelong Ao,Pengfei Lv,Jiancheng Dong,Di Wang,Yibing Cai,Qufu Wei,Yang Xu 성균관대학교(자연과학캠퍼스) 성균나노과학기술원 2018 NANO Vol.13 No.08

        Fluorescent nitrogen-doped carbon dots (N-CDs) with excellent stability were prepared via single-step hydrothermal carbonization of citric acid (CA) and ethylenediamine (EDA). The as-prepared N-CDs emit blue fluorescence under the excitation of 365 nm and have a size distribution of 2.80 ffi 0.47 nm with benign size effect. The structure and morphology were further characterized by X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) spectroscopy. It was found that the surface of the N-CDs was successfully functionalized, which presented water solubility and chelation with Fe3+. XRD results display a diffraction peak at 23.9 ℃, which corresponds to the (002) interlayer spacing of a graphitic structure revealing an amorphous carbon phase. Furthermore, due to good sensitivity, N-CDs were used as probes for Fe3+ detection. The low limit of detection of 0.6 μM as a fluorescence probe was successfully obtained based on the linear relationship between (F0 - F) / F0 and concentration of Fe3+ ions. Besides the satisfactory fluorescence, PVA/N-CDs membranes and fluorescent inks demonstrate potential for anti-counterfeiting applications due to its characteristic flexibility, transparency, removability and invisibility under ambient lighting.

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