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The effect of Zn(2+) on Pelodiscus sinensis creatine kinase: unfolding and aggregation studies.
Wang, Su-Fang,Lee, Jinhyuk,Wang, Wei,Si, Yue-Xiu,Li, Caiyan,Kim, Tae-Rae,Yang, Jun-Mo,Yin, Shang-Jun,Qian, Guo-Ying Adenine Press 2013 Journal of biomolecular structure & dynamics Vol.31 No.6
<P>We studied the effects of Zn(2+) on creatine kinase from the Chinese soft-shelled turtle, Pelodiscus sinensis (PSCK). Zn(2+) inactivated the activity of PSCK (IC(50)?=?.079??.004?mM) following first-order kinetics consistent with multiple phases. The spectrofluorimetry results showed that Zn(2+) induced significant tertiary structural changes of PSCK with exposure to hydrophobic surfaces and that Zn(2+) directly induced PSCK aggregation. The addition of osmolytes such as glycine, proline, and liquaemin successfully blocked PSCK aggregation, recovering the conformation and activity of PSCK. We measured the ORF gene sequence of PSCK by rapid amplification of cDNA end and simulated the 3D structure of PSCK. The results of molecular dynamics simulations showed that eight Zn(2+) bind to PSCK and one Zn(2+) is predicted to bind in a plausible active site of creatine and ATP. The interaction of Zn(2+) with the active site could mostly block the activity of PSCK. Our study provides important insight into the action of Zn(2+) on PSCK as well as more insights into the PSCK folding and ligand-binding mechanisms, which could provide important insight into the metabolic enzymes of P. sinensis.</P>
Alu Tandem Sequences Inhibit GFP Gene Expression by Triggering Chromatin Wrapping
Xiu Fang Wang,Xiao Yan Wang,Jing Liu,Jing Jing Feng,Wen Li Mu,Xiao Juan Shi,Qin Qing Yang,Xiao Cui Duan,Ying Xie,Zhan Jun Lu 한국유전학회 2009 Genes & Genomics Vol.31 No.3
Alu elements belonging to the short interspersed nuclear elements (SINE) of repetitive elements are present in more than one million copies which altogether represent 10% of the whole human genome. In this study, the roles of Alu tandem sequences in the process of GFP gene (GFP) expression and packing into chromatin of its DNA were studied. To detect the effect of Alu repeats on gene expression, different copies of Alus were inserted GFP downstream respectively in pEGFP-C1 vector. We found that Alu sequences decreased the amount of GFP transcription, the percentage of GFP positive cells and the accessibility to DNase I in length-dependent manner. Inserting Alu caused the production of higher-molecular-mass RNA, indicating Alu sequence did not induce premature transcriptional termination. Tight packing chromatins keep silent and resist to DNase I digestion, which is a general phenomenon. We suggested that head and tail tandem Alu sequences suppressed GFP expression in length dependent manner by triggering chromatin packing.
Wang, Li-Ying,Wang, Xiu-Hua,Tan, Jia-Lian,Xia, Shuai,Sun, Heng-Zhi,Shi, Jin-Wen,Jiang, Ming-Dong,Fang, Liang,Zuo, Hua,Dupati, Gautam,Jang, Kiwan,Shin, Dong-Soo Korean Chemical Society 2012 Bulletin of the Korean Chemical Society Vol.33 No.11
A number of novel small molecules, safrole oxide derivatives 4a-c, 6a-c, 9a-h, were synthesized by the reaction of safrole oxide with anilines 3 and 5, or its alkyl allyl ether derivative 7 with alkyl bromide 8 in moderate yields. The antiproliferative effects of all the target molecules on A549 cell growth were investigated and it was found that the 14 novel compounds could suppress A549 lung cancer cell growth. Among them, compound 6b was the most effective compound in inhibiting the proliferation of A549 cells.
Xiu-Shi Yang,Li-Li Wang,Xian-Rong Zhou,Shaomin Shuang,Zhi-Hua Zhu,Nan Li,Yan Li,Fang Liu,San-Cai Liu,Ping Lu,Guixing Ren,Chuan Dong 한국식품과학회 2013 Food Science and Biotechnology Vol.22 No.6
Quantitative detection of protein, fat, starch,and amino acids in foxtail millet using Fourier transformnear-infrared spectroscopy (NIRS) was investigated. Foxtail millet samples (n=259) were analyzed using NIRS. Spectral data were linearized with data from chemicalanalyses. Calibration models were established using apartial least-squares (PLS) algorithm with cross-validation. Optimized models were tested using external validation setsamples with coefficients of determination in the externalvalidation (R2val) of >0.90. Residual predictive deviation(RPD) values were nearly equal to or >2.5 for crudeprotein, alanine, aspartic acid, glutamic acid, isoleucine,leucine, and serine. However, for glycine, histidine,phenylalanine, proline, threonine, tyrosine, and valine, theR2val values were >0.83 and RPD values were nearly equalto or >2.0. For crude fat, total starch, arginine, and lysine,the R2val values were >0.70 and RPD values were >1.5. NIRS is a rapid determination tool for foxtail milletbreeding, and for quality control.
A New Cell Counting Method to Evaluate Anti-tumor Compound Activity
Wang, Xue-Jian,Zhang, Xiu-Rong,Zhang, Lei,Li, Qing-Hua,Wang, Lin,Shi, Li-Hong,Fang, Chun-Yan Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.8
Determining cell quantity is a common problem in cytology research and anti-tumor drug development. A simple and low-cost method was developed to determine monolayer and adherent-growth cell quantities. The cell nucleus is located in the cytoplasm, and is independent. Thus, the nucleus cannot make contact even if the cell density is heavy. This phenomenon is the foundation of accurate cell-nucleus recognition. The cell nucleus is easily recognizable in images after fluorescent staining because it is independent. A one-to-one relationship exists between the nucleus and the cell; therefore, this method can be used to determine the quantity of proliferating cells. Results indicated that the activity of the histone deacetylase inhibitor Z1 was effective after this method was used. The nude-mouse xenograft model also revealed the potent anti-tumor activity of Z1. This research presents a new anti-tumor-drug evaluation method.
Fang Jing,Jiang Xiao-Han,Wang Teng-Fei,Zhang Xiu-Jun,Zhang Ai-Di 한국식물생명공학회 2021 Plant biotechnology reports Vol.15 No.4
RNA editing is a significant post-transcriptional molecular process of modifying the primary transcripts by editosome. In plants, it remains unknown whether and to what extent RNA editing contributes to tissue-specific regulation from a global perspective. To obtain an overview of RNA editing events in model plant tobacco (Nicotiana tabacum), we implemented a bioinformatics analysis of DNA-Seq and RNA-Seq data from roots and leaves of three tobacco varieties (TN90, Basma and K326). The results showed that hundreds of RNA editing sites were detected to be located in the protein-coding region of plastid/mitochondria for all three varieties. Among these sites, some of them were detected in leaves but not or reduced in roots. Interestingly, most of the disappeared editing sites in roots were located in plastid transcripts encoding subunits of NADH dehydrogenase. The average editing efficiencies in roots were reduced significantly compared with leaves across three varieties in both organelles. In addition, we found that the reduction of RNA editing efficiency in mitochondria is mild compared with plastid. Expression analysis further showed that an extraordinarily high percentage of RNA editing factors were down-regulated in roots, particularly for PPR, MORF proteins, indicating that the distinct editing patterns between roots and leaves might result from the differential regulation of RNA editing factors expression. This study provides references and insights into understanding the function of plant RNA editing in tissue-specific regulation mediated by editosomes.
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.