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        D101 is critical for the function of AttJ, a repressor of quorum quenching system in Agrobacterium tumefaciens

        Chao Wang,Chunlan Yan,Yong-Gui Gao,Lian-Hui Zhang 한국미생물학회 2015 The journal of microbiology Vol.53 No.9

        The quorum quenching system of Agrobacterium tumefaciens is specifically activated upon entering the stationary phase. Evidence has shown that this system includes two key components: the IclR-type transcriptional factor AttJ (also named as BlcR) and the AHL-lactonase AttM (also named as BlcC). At exponential phase, AttJ binds to the promoter region of attM and thus suppresses the expression of attM. At stationary phase, however, the small molecule SSA directly binds to AttJ and relieves its inhibition of AttJ and thereby triggers the expression of attM. While the regulation of AttM has been extensively investigated, little is known about the regulation of AttJ. In this study, we demonstrated the D101 amino acid of AttJ is essential for the AttJ function. In vitro, the variant protein of AttJD101H appeared to be readily aggregated. In vivo, the D101H mutation in AttJ entirely abolished the inhibitory activity of AttJ and overexpressed attM in A. tumefaciens A6. In addition, D101H mutation led to an overexpression of attJ, indicating an auto-regulatory mechanism for the attJ regulation. Put together, these findings demonstrate that D101 is an important amino acid for the transcription activity of AttJ and the transcription of attJ is regulated by a negative feedback loop. These results expand previous biochemical characterization of AttJ and provide new mechanistic insights into the regulation of quorum quenching in A. tumefaciens.

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        Synthesis of Frataxin Genes by Direct Assembly of Serial Deoxyoligonucleotide Primers and Its Expression in Escherichia coli

        윤영걸,Sun-Hee Park,Jee Suk Lee,Chunlan Yan,Chan Bae Park,Michael D Koob,유영현 한국생물공학회 2013 Biotechnology and Bioprocess Engineering Vol.18 No.2

        Frataxin, a small nuclear-encoded protein targeted to mitochondria, is known to play an important role in both the mitochondrial respiratory chain and iron homeostasis. The protein is highly conserved in most eukaryotic organisms with no major structural changes, suggesting that it serves a crucial function in all organisms. Recently,purified frataxin was used as a therapeutic treatment of Friedreich’s ataxia, a common degenerative disorder that results from a frataxin protein deficiency, by directly applying the protein to the diseased cells. In this report, we describe a novel and rapid method of synthesizing genes encoding frataxin proteins for the purpose of efficient protein production. The artificial yeast and human frataxin genes were synthesized by direct assembly of serial deoxyoligonucleotide primers designed based on the optimal nucleotide sequences. When we tested the expression of these synthetic genes in two E. coli host strains, the yeast frataxin gene was expressed 20 folds higher in Rosetta (DE3) cells than in BL21 (DE3) cells, whereas the expression levels of human frataxin were similar in both E. coli strains. Attenuation of the Fenton reactions by the purified yeast and human frataxin proteins was observed under the defined conditions, which suggests that the recombinant frataxin proteins are active and functional. The procedure described here could be applied to many known genes or to generate novel synthetic genes that can be redesigned by arranging functional domains from previously identified genes and to study the structure and function of synthetic recombinant proteins and potential usage.

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        Scaling analysis of magnetic-thermal behaviors in ferromagnetic insulator LaCoO3 thin film

        Liu Hao,Fan Jiyu,Zheng Huan,Rahman Azizur,Qian Fengjiao,Wang Caixia,Ma Chunlan,Zhu Yan,Yang Hao 한국물리학회 2021 Current Applied Physics Vol.28 No.-

        The structure, surface topography and magnetic properties of LaCoO3 (LCO) thin films deposited epitaxially on the SrTiO3 substrates have been investigated in detail. The LCO thin films show a typical ferromagneticparamagnetic (FM-PM) phase transition at 78 K. Based on the measurements of isothermal magnetization around the Curie point, the magnetic entropy change (ΔSM) of LCO thin films under various applied magnetic fields were obtained. By utilizing scaling theory, all of the ΔSM curves can be re-scaled, confirming that the FMPM phase transition is second-order magnetic phase transition. Moreover, the magnetic entropy change -ΔSM features a maximum around TC, whereas, power law fitting of -ΔSmax M with H gives n = 0.9704, which is obviously deviating from the standard value n = 2/3 for the Landau mean field model. This indicates that the mean field theory cannot be used to explain the critical behavior of LCO thin films which implies that the critical behavior of LCO thin films may involve complex magnetic interactions. These interactions are correlated with a long-term puzzling finding in this system why its Curie point is always around 80 K regardless of the different fabrication methods and conditions used in the film growth.

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