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A library of TAL effector nucleases spanning the human genome
Kim, Yongsub,Kweon, Jiyeon,Kim, Annie,Chon, Jae Kyung,Yoo, Ji Yeon,Kim, Hye Joo,Kim, Sojung,Lee, Choongil,Jeong, Euihwan,Chung, Eugene,Kim, Doyoung,Lee, Mi Seon,Go, Eun Mi,Song, Hye Jung,Kim, Hwangbeo Nature Publishing Group, a division of Macmillan P 2013 Nature biotechnology Vol.31 No.3
Transcription activator–like (TAL) effector nucleases (TALENs) can be readily engineered to bind specific genomic loci, enabling the introduction of precise genetic modifications such as gene knockouts and additions. Here we present a genome-scale collection of TALENs for efficient and scalable gene targeting in human cells. We chose target sites that did not have highly similar sequences elsewhere in the genome to avoid off-target mutations and assembled TALEN plasmids for 18,740 protein-coding genes using a high-throughput Golden-Gate cloning system. A pilot test involving 124 genes showed that all TALENs were active and disrupted their target genes at high frequencies, although two of these TALENs became active only after their target sites were partially demethylated using an inhibitor of DNA methyltransferase. We used our TALEN library to generate single- and double-gene-knockout cells in which NF-κB signaling pathways were disrupted. Compared with cells treated with short interfering RNAs, these cells showed unambiguous suppression of signal transduction.
Han, Songhee,Hanh Nguyen, Thi Thanh,Hur, Jaewon,Kim, Nahyun M.,Kim, Seong-Bo,Hwang, Kyeong-Hwan,Moon, Young-Hwan,Kang, Choongil,Chung, Byoungsang,Kim, Young-Min,Kim, Tae Sung,Park, Jun-Seong,Kim, Doma Elsevier 2017 Enzyme and microbial technology Vol.103 No.-
<P>Astragalin (kaempferol-3-O-beta-D-glucopyranoside, Ast) is a kind of flavonoid known to have anti-oxidant, anti-HIV, anti-allergic, and anti-inflammatory effects. It has low solubility in water. In this study, novel astragalin galactosides (Ast-Gals) were synthesized using beta-galactosidase from Bacillus circulans and reaction conditions were optimized to increase the conversion yield of astragallin. Purified Ast-Gal1 (11.6% of Ast used, w/w) and Ast-Gal2 (6.7% of Ast used, w/w) were obtained by medium pressure chromatography (MPLC) with silica C-18 column and open column packed with Sephadex LH-20. The structures of Ast-Gal1 and Ast-Gal2 were identified by nuclear magnetic resonance (NMR) to be kaempferol-3-O-beta-D-glucopyranosyl-(1 -> 6)-beta-D-galactopyranoside and kaempferol-3-O-beta-D-glucopyranosyl-(1 -> 6)-beta-D-galactopyranosyl-(1 -> 4)-beta-D-galactopyranoside, respectively. The water solubility of Ast, Ast-Gal1, and Ast-Gal2 were 28.2 +/- 1.2 mg/L, 38,300 +/- 3.5 mg/L, and 38,800 +/- 2.8 mg/L, respectively. The SC50 value (the concentration required to scavenge 50% of the ABTS.+) of Ast, Ast-Gal1, and Ast-Gal2 were 5.1 +/- 1.6 mu M, 6.5 +/- 0.4 mu M, and 4.9 +/- 1.1 mu M, respectively. The IC50 values (the half maximal inhibitory concentration) of Ast, Ast-Gal1, and Ast-Gal2 against angiotensin converting enzyme (ACE) were 171.0 +/- 1.2 mu M, 186.0 mu M, and 139.0 +/- 0.2 mu M, respectively.</P>
컴퓨터 기반 및 과정중심의 작문교육에 있어서 상호작용에 대한 연구
김충일(Kim Choongil) 팬코리아영어교육학회(구 영남영어교육학회) 2005 영어교육연구 Vol.17 No.4
The purpose of this study is to find plausible differences in the quality of writing products across the instructional modes with and without interaction in writing, and students’ attitudes toward interaction, the computer use, and the process-based approach. Thirty eight Korean EFL high school students produced three essays under the CALL-based and process- based instruction with the experiment group having the treatment of interaction. Before and after the course, the timed pre- and post-tests were given. Out of 5 essays, the pre-test, the post-test, and the final draft of the second cycle were scored holistically and analytically. In addition, after every class in the first cycle, written interviews were administered to find out the students’ attitudes about what they had gone through during the classes. After the course, a brief survey for finding their general feelings about the main issues of this study was conducted. According to the results, the scores for the pre-test and the post-test showed no statistical difference; however, the experiment’s scores for the final draft of the second cycle were significantly higher than those of the control. The process-based approach, the computer use, and interaction were considered favorable and helpful by both groups according to the results of written interviews and the post survey with the former being more positive.
Efficient PRNP deletion in bovine genome using gene-editing technologies in bovine cells.
Choi, WooJae,Kim, Eunji,Yum, Soo-Young,Lee, ChoongIl,Lee, JiHyun,Moon, JoonHo,Ramachandra, Sisitha,Malaweera, Buddika Oshadi,Cho, JongKi,Kim, Jin-Soo,Kim, SeokJoong,Jang, Goo Landes Bioscience 2015 Prion Vol.9 No.4
<P>Even though prion (encoded by the PRNP gene) diseases like bovine spongiform encephalopathy (BSE) are fatal neurodegenerative diseases in cattle, their study via gene deletion has been limited due to the absence of cell lines or mutant models. In this study, we aim to develop an immortalized fibroblast cell line in which genome-engineering technology can be readily applied to create gene-modified clones for studies. To this end, this study is designed to 1) investigate the induction of primary fibroblasts to immortalization by introducing Bmi-1 and hTert genes; 2) investigate the disruption of the PRNP in those cells; and 3) evaluate the gene expression and embryonic development using knockout (KO) cell lines. Primary cells from a male neonate were immortalized with Bmi-1and hTert. Immortalized cells were cultured for more than 180??days without any changes in their doubling time and morphology. Furthermore, to knockout the PRNP gene, plasmids that encode transcription activator-like effector nuclease (TALEN) pairs were transfected into the cells, and transfected single cells were propagated. Mutated clonal cell lines were confirmed by T7 endonuclease I assay and sequencing. Four knockout cell lines were used for somatic cell nuclear transfer (SCNT), and the resulting embryos were developed to the blastocyst stage. The genes (CSNK2A1, FAM64A, MPG and PRND) were affected after PRNP disruption in immortalized cells. In conclusion, we established immortalized cattle fibroblasts using Bmi-1 and hTert genes, and used TALENs to knockout the PRNP gene in these immortalized cells. The efficient PRNP KO is expected to be a useful technology to develop our understanding of in vitro prion protein functions in cattle.</P>
김창대(Changdae Kim),권충일(Choongil Kwon),김석재(Seokjae Kim),여권구(Gwonkoo Yeo) 한국자동차공학회 2002 한국자동차공학회 춘 추계 학술대회 논문집 Vol.2002 No.5_1
To meet future EURO-III, IV emission standards, it is essential to develop catalyst technology that will reduce diesel engine exhaust gases. In this study, deactivation of diesel oxidation catalyst used in fleet vehicle test was mainly investigated To conform the surface characterization of catalyst, BET, IC & CO chemisorption were used.<br/> The causes of deactivation of a diesel catalyst were several but the main factors were sulfur poisoning, carbon coking & Pt sintering among them. Therefore, the LOT of an endurance sample was increased higher than thai of a fresh Especially, the change of pore diameter & volume by sulfur was observed.
Koo, Ok Jae,Park, Sol Ji,Lee, Choongil,Kang, Jung Taek,Kim, Sujin,Moon, Joon Ho,Choi, Ji Yei,Kim, Hyojin,Jang, Goo,Kim, Jin-Soo,Kim, Seokjoong,Lee, Byeong-Chun Asian Australasian Association of Animal Productio 2014 Animal Bioscience Vol.27 No.3
To facilitate the construction of genetically-modified pigs, we produced cloned embryos derived from porcine fibroblasts transfected with a pair of engineered zinc finger nuclease (ZFN) plasmids to create targeted mutations and enriched using a reporter plasmid system. The reporter expresses RFP and eGFP simultaneously when ZFN-mediated site-specific mutations occur. Thus, double positive cells ($RFP^+/eGFP^+$) were selected and used for somatic cell nuclear transfer. Two types of reporter based enrichment systems were used in this study; the cloned embryos derived from cells enriched using a magnetic sorting-based system showed better developmental competence than did those derived from cells enriched by flow cytometry. Mutated sequences, such as insertions, deletions, or substitutions, together with the wild-type sequence, were found in the cloned porcine blastocysts. Therefore, genetic mutations can be achieved in cloned porcine embryos reconstructed with ZFN-treated cells that were enriched by a reporter-based system.
하향링크 다중 사용자 다중 입출력 채널을 위한 향상된 Block Tomlinson-Harashima Precoding 기법
김준두(Joon-doo Kim),강지원(Jiwon Kang),최인경(Inkyeong Choi),예충일(Choongil Yeh),권동승(Donseung Kwon),이충용(Chungyong Lee) 대한전자공학회 2007 대한전자공학회 학술대회 Vol.2007 No.11
In this thesis, we propose two algorithms which improve the error performance of the BTHP system. The proposed expanded ML (eML) algorithm provides an approach to apply the ML receiver to the BTHP system. The proposed block QR (bQR) algorithm makes the effective MIMO channel that the proposed eML can obtain spatial diversity gain from it. By applying the proposed methods, BTHP obtains the spatial diversity and considerable error performance improvement.
Chin, Chong Shik,Eum, Min-Sik,Kim, Song yi,Kim, Choongil,Kang, Sung Kwon WILEY-VCH Verlag 2006 European journal of inorganic chemistry Vol.2006 No.24
<P>A new type of iridium(III) complex [trans-Ir(ppy)<SUB>2</SUB>(PPh<SUB>3</SUB>)<SUB>2</SUB>]<SUP>+</SUP> (1) has been prepared by a novel synthetic method and its structural and photoluminescent characteristics have been compared with those of the cis analogue, [cis-Ir(ppy)<SUB>2</SUB>(PPh<SUB>3</SUB>)(P(OPh)<SUB>3</SUB>)]<SUP>+</SUP> (2) which has also been newly prepared in this study. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2006)</P> <B>Graphic Abstract</B> <P> <img src='wiley_img/14341948-2006-2006-24-EJIC200600888-fig000.gif' alt='wiley_img/14341948-2006-2006-24-EJIC200600888-fig000'> </P>
Subburaj, Saminathan,Chung, Sung Jin,Lee, Choongil,Ryu, Seuk-Min,Kim, Duk Hyoung,Kim, Jin-Soo,Bae, Sangsu,Lee, Geung-Joo Springer International 2016 Plant cell reports Vol. No.
<P>Site-directed mutagenesis of nitrate reductase genes using direct delivery of purified Cas9 protein preassembled with guide RNA produces mutations efficiently in Petunia x hybrida protoplast system. The clustered, regularly interspaced, short palindromic repeat (CRISPR)-CRISPR associated endonuclease 9 (CRISPR/Cas9) system has been recently announced as a powerful molecular breeding tool for site-directed mutagenesis in higher plants. Here, we report a site-directed mutagenesis method targeting Petunia nitrate reductase (NR) gene locus. This method could create mutations efficiently using direct delivery of purified Cas9 protein and single guide RNA (sgRNA) into protoplast cells. After transient introduction of RNA-guided endonuclease (RGEN) ribonucleoproteins (RNPs) with different sgRNAs targeting NR genes, mutagenesis at the targeted loci was detected by T7E1 assay and confirmed by targeted deep sequencing. T7E1 assay showed that RGEN RNPs induced site-specific mutations at frequencies ranging from 2.4 to 21 % at four different sites (NR1, 2, 4 and 6) in the PhNR gene locus with average mutation efficiency of 14.9 +/- 2.2 %. Targeted deep DNA sequencing revealed mutation rates of 5.3-17.8 % with average mutation rate of 11.5 +/- 2 % at the same NR gene target sites in DNA fragments of analyzed protoplast transfectants. Further analysis from targeted deep sequencing showed that the average ratio of deletion to insertion produced collectively by the four NR-RGEN target sites (NR1, 2, 4, and 6) was about 63:37. Our results demonstrated that direct delivery of RGEN RNPs into protoplast cells of Petunia can be exploited as an efficient tool for site-directed mutagenesis of genes or genome editing in plant systems.</P>
신종식,박정한,김충일,Chin Chong Shik,Park Jeonghan,Kim Choongil Korean Chemical Society 1989 Bulletin of the Korean Chemical Society Vol.10 No.1
Four coordinated rhodium(Ⅰ) complexes, Rh($ClO_4$)(CO)$(PPh_3)_2$ and [$Rh(CO)(PPh_3)_3$]$ClO_4$(2) catalyze the iosmerization of allylic alcohols to the corresponding carbonyl compounds at room temperature under nitrogen. The isomerization is faster with 2 than with 1, which is understood in terms of relative ease of the last step of the catalytic cycle, the reductive elimination of enol. Relative rates of the isomerization with 1 and 2 for different allylic alcohols are also explained by the relative ease of the enol elimination step in part. The first step of the catalytic cycle, the complex formation of the allylic alcohol through the ${\pi}-system$ of the olefinic group of the allylic alcohol and the following step, formation of hydridoallyl complex also seem to affect the overall rate of the isomerization.