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Cloning and Expression of Mycobacterium bovis Secreted Protein MPB83 in Escherichia coli
Xiu-Yun, Jiang,Wang, Chun-Feng,Wang, Chun-Fang,Zhang, Peng-Ju,He, Zhao-Yang Korean Society for Biochemistry and Molecular Biol 2006 Journal of biochemistry and molecular biology Vol.39 No.1
The gene encoding MPB83 from Mycobacterium bovis Vallee111 chromosomal DNA was amplified by using polymerase chain reaction (PCR) technique, and the PCR product was approximately 600bp DNA segment. Using T-A cloning technique, the PCR product was cloned into pGEM-T vector and the cloning plasmid pGEM-T-83 was constructed successfully. pGEM-T-83 and pET28a(+) were digested by BamHI and EcoRI double enzymes. The purified MPB83 gene was subcloned into the expression vector pET28a(+), and the prokaryotic expression vector pET28a-83 was constructed. Plasmid containing pET28a-83 was transformed into competence Escherichia coli BL21 (DE3). The bacterium was induced by isopropyl-$\beta$-D-thiogalactopyranoside (IPTG) and its lysates were loaded directly onto sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE), approximately 26 kDa exogenous protein was observed on the SDS-PAGE. The protein was analyzed using Western-blotting. The results indicated that the protein was of antigenic activity of M. bovis. The results were expected to lay foundation for further studies on the subunit vaccine and DNA vaccine of MPB83 gene in their prevention against bovine tuberculosis.
Ultrafast UV Switch Based on ZnO-Ag Heterostructures
XIU-YUN AN,Feng Teng,Zhenxing Zhang,Xiaojun Pan,Jinyuan Zhou,ER-QING XIE 대한금속·재료학회 2014 ELECTRONIC MATERIALS LETTERS Vol.10 No.1
ZnO-silver (Ag) heterostructure nanoparticle films were prepared by spin-coating, followed by annealing at 700°C for 2 h. The films were then used as UV photodetector which show high photoresponse. The heterostructure-film device displayed an ultrafast decay time of 18 ms and a rise time of 50 ms upon ultraviolet irradiation. Additionally, the time-dependent photocurrent upon UV switching reveals a rectangularly shaped profile, rarely reported in previous literature. The highly improved photoresponse properties of ZnO-Ag heterostructure-film device could be attributed to the Schottky barrier height (SBH) and depletion width reduction from the embedded Ag nanoclusters. Compared to a pure ZnO film, both the responsivity (Rλ) and external quantum efficiency (EQE) of the ZnO-Ag heterostructure-film photodetectors were improved more than 13-fold. This research provides a promising strategy for fabricating UV-photodetectors with ultrafast response.
Highly selective two-photon imaging of cysteine in cancerous cells and tissues
Lee, Yun Hak,Ren, Wen Xiu,Han, Jiyou,Sunwoo, Kyoung,Lim, Ja-Yun,Kim, Jong-Hoon,Kim, Jong Seung The Royal Society of Chemistry 2015 Chemical communications Vol.51 No.76
<P>Abnormal concentrations of Cys have been reported to be implicated in various health problems, including cancer, neuropathy, and cardiomyopathy. We present a novel two-photon fluorescent probe for the specific recognition of cysteine over homocysteine and glutathione, and the bioapplication of this probe for the imaging of live cancerous cells and thick tissues.</P> <P>Graphic Abstract</P><P>Abnormal concentrations of Cys have been reported to be implicated in various health problems, including cancer, neuropathy, and cardiomyopathy. <IMG SRC='http://pubs.rsc.org/services/images/RSCpubs.ePlatform.Service.FreeContent.ImageService.svc/ImageService/image/GA?id=c5cc06038a'> </P>
Zhan, Yun-Hong,Liu, Jing,Qu, Xiu-Juan,Hou, Ke-Zuo,Wang, Ke-Feng,Liu, Yun-Peng,Wu, Bin Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.6
Background: Renal-cell carcinoma (RCC) is resistant to almost all chemotherapeutics and radiation therapy. ${\beta}$-Elemene, a promising anticancer drug extracted from a traditional Chinese medicine, has been shown to be effective against various tumors. In the present study, anti-tumor effects on RCC cells and the involved mechanisms were investigated. Methods: Human RCC 786-0 cells were treated with different concentrations of ${\beta}$-elemene, and cell viability and apoptosis were measured by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay and flow cytometry, respectively. Protein expression was assayed by western blotting. Autophagy was evaluated by transmission electron microscopy. Results: ${\beta}$-Elemene inhibited the viability of 786-0 cells in a dose- and time-dependent manner. The anti-tumor effect was associated with induction of apoptosis. Further study showed that ${\beta}$-elemene inhibited the MAPK/ERK as well as PI3K/Akt/mTOR signalling pathways. Moreover, robust autophagy was observed in cells treated with ${\beta}$-elemene. Combined treatment of ${\beta}$-elemene with autophagy inhibitors 3-methyladenine or chlorochine significantly enhanced the anti-tumor effects. Conclusions: Our data provide first evidence that ${\beta}$-elemene can inhibit the proliferation of RCC 786-0 cells by inducing apoptosis as well as protective autophagy. The anti-tumor effect was associated with the inhibition of MAPK/ERK and PI3K/Akt/mTOR signalling pathway. Inhibition of autophagy might be a useful way to enhance the anti-tumor effect of ${\beta}$-elemene on 786-0 cells.
( Xiao Yun Huang ),( Juan Lin ),( Xiu Yun Ye ),( Guo Zeng Wang ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.5
To enrich the genetic resource of microbial xylanases with high activity and stability under alkaline conditions, a xylanase gene (xynSL4) was cloned from Planococcus sp. SL4, an alkaline xylanase-producing strain isolated from the sediment of soda lake Dabusu. Deduced XynSL4 consists of a putative signal peptide of 29 residues and a catalytic domain (30-380 residues) of glycosyl hydrolase family 10, and shares the highest identity of 77% with a hypothetical protein from Planomicrobium glaciei CHR43. Phylogenetic analysis indicated that deduced XynSL4 is closely related with thermophilic and alkaline xylanases from Geobacillus and Bacillus species. The gene xynSL4 was expressed heterologously in Escherichia coli and the recombinant enzyme showed some superior properties. Purified recombinant XynSL4 (rXynSL4) was highly active and stable over the neutral and alkaline pH range from 6 to 11, with maximum activity at pH 7 and more than 60% activity at pH 11. It had an apparent temperature optimum of 70oC and retained stable at this temperature in the presence of substrate. rXynSL4 was highly halotolerant, retaining more than 55% activity with 0.25.3.0 M NaCl and was stable at the concentration of NaCl up to 4M. The enzyme activity was significantly enhanced by β-mercaptoethanol and Ca2+ but strongly inhibited by heavy-metal ions and SDS. This thermophilic and alkaline- and salt-tolerant enzyme has great potential for basic research and industrial applications.
( Yun Deng ),( Bi Sheng Liu ),( Xiong Wei Fan ),( Yue Qun Wang ),( Ming Tang ),( Xiao Yang Mo ),( Yong Qing Li ),( Zao Chu Ying ),( Yong Qi Wan ),( Na Luo ),( Jun Mei Zhou ),( Xiu Shan Wu ),( Wu Zhou 한국생화학분자생물학회 (구 한국생화학회) 2010 BMB Reports Vol.43 No.3
In this study, we report the identification and characterization of a novel C2H2 zinc finger protein, ZNF552, from a human embryonic heart cDNA library. ZNF552 is composed of three exons and two introns and maps to chromosome 19q13.43. The cDNA of ZNF552 is 2.3 kb, encoding 407 amino acids with an amino-terminal KRAB domain and seven carboxyl-terminal C2H2 zinc finger motifs in the nucleus and cytoplasm. Northern blotting analysis indicated that a 2.3 kb transcript specific for ZNF552 was expressed in liver, lung, spleen, testis and kidney, especially with a higher level in the lung and testis in human adult tissues. Reporter gene assays showed that ZNF552 was a transcriptional repressor, and overexpression of ZNF552 in the COS-7 cells inhibited the transcriptional activities of AP-1 and SRE, which could be relieved through RNAi analysis. Deletion studies showed that the KRAB domain of ZNF552 may be involved in this inhibition. [BMB reports 2010; 43(3): 193-198]
A Role of YlBud8 in the Regulation of Cell Separation in the Yeast Yarrowia lipolytica
( Yun-qing Li ),( Qing-jie Xue ),( Yuan-yuan Yang ),( Hui Wang ),( Xiu-zhen Li ) 한국미생물 · 생명공학회 2019 Journal of microbiology and biotechnology Vol.29 No.1
The spatial landmark protein Bud8 plays a crucial role in bipolar budding in the budding yeast Saccharomyces cerevisiae. The unconventional yeast Yarrowia lipolytica can also bud in a bipolar pattern, but is evolutionarily distant from S. cerevisiae. It encodes the protein YALI0F12738p, which shares the highest amino acid sequence homology with S. cerevisiae Bud8, sharing a conserved transmembrane domain at the C-terminus. Therefore, we named it YlBud8. Deletion of YlBud8 in Y. lipolytica causes cellular separation defects, resulting in budded cells remaining linked with one another as cell chains or multiple buds from a single cell, which suggests that YlBud8 may play an important role in cell separation, which is distinct from the function of Bud8 in S. cerevisiae. We also show that the YlBud8-GFP fusion protein is located at the cell membrane and enriched in the bud cortex, which would be consistent with a role in the regulation of cell separation. The coiled-coil domain at the Nterminus of YlBud8 is important to the correct localization and function of YlBud8, as truncated proteins that do not contain the coiled-coil domain cannot rescue the defects observed in Ylbud8Δ. This finding suggests that a new signaling pathway controlled by YlBud8 via regulation of cell separation may exist in Y. lipolytica.