• Title/Summary/Keyword: 분자 계통분석

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Inference of Gene Phylogenetic Tree based on Decision Tree (결정트리 분류기법 기반 유전자 계통수 추론)

  • 김신석;황부현
    • Proceedings of the Korean Information Science Society Conference
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    • 2001.10a
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    • pp.280-282
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    • 2001
  • 분자생물학의 급진적 발전은 현대 계통분류학에 큰 변혁을 가져왔다. 특히 유전의 근원물질인 DNA나 RNA를 분리.조작.분석하는 기술의 발전으로 이를 이용만 계통수 제작은 계통생물학의 중요한 실험방법으로 자리잡고 있다. 그 중 염기서열 비교 방법은 현재 유전자 계통수 제작에 가장 널리 이용되는 방법이다. 하지만 이러만 계통수는 각 객체간의 거리만을 표현하고, 객체군간의 차이는 설명하기 힘들다. 본 연구에서는 염기서열의 상대적인 특징(유사도)을 대신하는 염기서열의 총량과 염기 함량 등을 이용해 새로이 분류 기법 중 결정트리 방법에 적응하고, 종 분류의 유전적 모델을 설계한다. 또한 결정트리의 클래스인 종은 상위 클래스들을 포함하고 있어, 본 논문에서는 기존의 결정트리 분류자를 수정한 단계적 결정트기 분류자를 제안한다.

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Development of salt-tolerant transgenic chrysanthemum (Dendranthema grandiflorum) lines and bio-assay with a change of cell specificity (내염성 국화 형질전환 계통 육성 및 저항성 검정과 세포특성 변화)

  • Kang, Chan-Ho;Yun, Seung-Jung;Han, Bum-So;Lee, Gong-Joon;Choi, Kyu-Hwan;Park, Jong-Suk;Shin, Yong-Kyu
    • Journal of Plant Biotechnology
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    • v.38 no.1
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    • pp.1-8
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    • 2011
  • Recently the increasing of vinyl and green houses and development of reclaimed land including Saemangeum induced the need for breeding salt-tolerant crops which can survive and grow in high salinity soil. So we try to develop salt-tolerant transgenic chrysanthemum (Dendranthema grandiflorum.) lines by using anti-porter gene TANHX and HVNHX. Through marker selection and plant regeneration step, we could get 284 putative transgenic chrysanthemum lines. On selected putative transgenic plants, 40 candidates were used for genetic analysis and 30 lines could be made up of target size band on PCR, so about 75% of marker selected lines were decided as real transgenic lines. Selected 284 transgenic lines were also used for salt-tolerance test as a range of NaCl 0.2 ~ 1.2% (300 mM). As a result of salt-tolerance test, 15 selected transgenic lines could live and grow on the continuous supply of 0.8% (200 mM) NaCl solution and another 7 lines were could survive under 1.2% (300 mM) NaCl solution. This salt-tolerant transgenic lines under salt stress also lead a cell alternation especially a guard cell. A stressed guard cell be swelled and grow larger in proportion to NaCl concentration. TTC test for cell viability on transgenic chrysanthemum lines pointed out that more strong salt-tolerant lines can be live more than another under same salt stress. The numerical value of strong salt-tolerant 7 transgenic lines were 0.206 ~ 0.331 under 1.2% NaCl stress, and then it's value is more larger than middle salinity lines' 0.114 ~ 0.193 and non-transgenic's 0.046. And the proline contents as indicated stress compound also pointed out that HVNHX introduced salt-tolerant transgenic lines were less stressed than other under same salt stress. The contents of strong salt-tolerant transgenic lines were 2.255 ~ 2.638 mg/kg and it is much higher than that of middle salinity lines' 1.496 ~ 2.125.

Molecular Characterization and Event-Specific Marker Development of Insect Resistant Chinese Cabbage for Environmental Risk Assessment (환경위해성 평가를 위한 해충저항성 배추의 분자생물학적 특성 검정 및 계통 특이 마커 캐발)

  • Lim, Sun-Hyung;Kim, Na-Young;Lee, Si-Myung;Woo, Hee-Jong;Shin, Kong-Sik;Jin, Yong-Moon;Cho, Hyun-Suk
    • Journal of Plant Biotechnology
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    • v.34 no.4
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    • pp.347-354
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    • 2007
  • Commercialization of genetically modified (GM) plants will be required the assessment of risks associated with the release of GM plants that should include a detailed risk assessment of their impacts in human health and the environment. Prior to GM plant release, applicants should provide the information on GM crops for approval. We carried out this study to provide the molecular data for risk assessment of the GM Chinese cabbage plants with insect-resistance gene, modified CryIAc, which we obtained by Agrobacterium-transformation. From the molecular analysis with GM Chinese cabbage, we confirmed the transgene copy number and stability, the expression of the transgene, and integration region sequences between the transgene and the Chinese cabbage genome. Based on the unique integration DNA sequences, we designed specific primer set to detect GM Chinese cabbage and set up the GM cabbage detection method by qualitative PCR analysis. Qualitative analysis with GM Chinese cabbage progenies analysis was revealed the same as the result of herbicide treatment. Our results provided the molecular data for risk assessment analysis of GM Chinese cabbage and demonstrated that the primer set proposed could be useful to detect GM Chinese cabbage.

Molecular Characterization of a Korean Isolate of Human Norovirus, the Hu/NLV/Gunpo/2006/KO Strain (인체 노로바이러스의 한국분리주 Hu/NLV/Gunpo/2006/KO의 분자생물학적 특성)

  • Jeong, Ah-Yong;Yun, Sang-Im;Jee, Young-Mee;Kang, Yoon-Sung;Lee, Young-Min
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.105-111
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    • 2009
  • Norovirus (NV) with a variety of genotypes, a member of the family Caliciviridae, causes acute nonbacterial gastroenteritis in humans. We determined the nucleotide sequence of three open reading frames (ORFs) of a NV Korean strain and characterized the genetic relationship with others. The Korean strain designated Hu/NLV/Gunpo/2006/KO was isolated from the stool specimen of a 2-year-old female suffering from gastroenteritis. By performing reverse transcription and PCR amplification, three overlapping cDNAs were synthesized and used for direct sequencing. We found that like other NVs, this strain contains three ORFs: ORF1, 5,100 bp; ORF2, 1,647 bp; ORF3, 765 bp. Of 35 NVs, ORF1 had a level of genetic diversity lower than ORF2 and ORF3, of which the C-termini of the ORF2 and ORF3 showed a relatively high degree of genetic diversity. Phylogenetic analyses indicated that the Korean strain belonged to genogroup II, with Saitama U1, Gifu'96, Mc37, and Vietnam 026 being formed a single genetic cluster. The nucleotide sequence information of three ORFs of a NV Korean isolate will be useful not only for the development of a diagnostic tool and understanding of genetic relationship, but also provide important basic information for the functional analysis of their gene products.

Identification of Korean Mountain Cultivated Ginseng by RAPD (RAPD(Random Amplified Polymorphic DNA)를 이용한 장뇌삼의 지역별 품종 구분)

  • Choi, Ji-Young;Lee, Ju-Hee;Lee, Su-Gwang;Kang, Ho-Duck
    • Journal of agriculture & life science
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    • v.43 no.6
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    • pp.35-43
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    • 2009
  • This study was conducted to examine the genetic variations and intraspecific relationships between 9 individuals of Panax ginseng C.A Meyer by using RAPD (Randomly Amplified Polymorphic DNA) analysis. The 34 primers out of 40 random primers were amplified for all tested plants. The 48 (40%) among 244 bands derived from 34 primers shown polymorphism, and the 72 (64%) rest of bands showed similar forms. By regional groups Sangju and Andong samples located in Kyungsang buk-do showed a high similarity. However, Punggi located in Kyungsang buk-do showed higher similarity with Jinan's of Junla buk-do. In this way, it did not show that Panax ginseng from the same area has similarities. In future study we need to more specific molecular phylogenetic analysis such as AFLP technology and gene sequencing with nuclear chloroplast DNA in all samples.

Development of Near-Isogenic Line of japonica Rice Cultivar Saenuri without Lipoxygenase-3 (새누리 벼 품종 배경 lipoxygenase-3 결핍 자포니카 근동질계통 개발)

  • Park, Hyun-Su;Lee, Keon-Mi;Kim, Ki-Young;Kim, Jeong-Ju;Shin, Woon-Cheol;Baek, Man-Kee;Kim, Choon-Song;Park, Seul-Gi;Lee, Chang-Min;Suh, Jung-Pil;Cho, Young-Chan
    • Korean Journal of Breeding Science
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    • v.51 no.3
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    • pp.190-200
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    • 2019
  • It is reported that the absence of lipoxygenase-3 (LOX-3) may contribute to a reduction in stale flavor after the storage of rice. To improve the quality of stored rice of the Korean japonica rice cultivar, we conducted a breeding program to develop near-isogenic rice without LOX-3 in the genetic background of Saenuri, a mega variety of Korea. In the first step of the breeding program, we used a donor parent of LOX-3 null, Daw Dam, and a recurrent japonica parent, Sindongjin, to develop HR27873-AC12 by backcross (BC1), color test for introgression of lox-3, and anther culture for rapid fixation. In the second step, we used the donor parent, HR27873-AC12, and the recurrent parent, Saenuri, to develop HR28896-31-3-1-1 by backcross (BC1), marker-assisted selection (MAS) for lox-3, and phenotypic selection (PS) for agronomic traits. Finally, in the third step, we developed HR30960-186-2-1-2-1 (Jeonju624), derived from a cross between Saenuri and HR28896-31-3-1-1, by MAS for lox-3 and PS with high selection pressure for agronomic characteristics. Jeonju624 was confirmed with the introgression of lox-3 by molecular marker. Jeonju624 was a mid-late maturing rice with similar agronomic characteristics to Saenuri, lodging tolerance with short culm, erect plant architecture, and resistance to bacterial blight and rice stripe virus. The yield components of Jeonju624 were mostly similar to Saenuri, except for the 1,000-grain weight of brown rice. The appearance of the grain of Jeonju624 was better than that of Saenuri, and the characteristics of cooked rice were similar to those of Saenuri. In the genetic background analysis using 406 KASP (Kompetitive Allele-Specific PCR) markers, Jeonju624 was confirmed to be the near-isogenic line (NIL) of Saenuri with a 95.8% recovery rate. Jeonju624 is the NIL of Saenuri without LOX-3, and overcomes the linkage drag of Daw Dam with similar agronomic characteristics and genetic background to Saenuri. Jeonju624 can be utilized as a practical cultivar to improve the quality of stored rice, breeding material for the introgression of lox-3, and genetic material to elucidate the effect of introgressed genes.

Studies on Genetic Diversity and Phylogenetic Relationships of Korean Native Chicken using the Microsatellite Marker (Microsatellite Marker를 활용한 한국 토종닭 품종의 유전적 다양성 및 유연관계 분석)

  • Seo, Joo Hee;Oh, Jea-Don;Lee, Jun-Heon;Seo, Dongwon;Kong, Hong Sik
    • Korean Journal of Poultry Science
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    • v.42 no.1
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    • pp.15-26
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    • 2015
  • In this study, genotyping was executed by using 27 microsatellite markers for genetic diversity of 469 Korean Native Chickens [20 population, each population is 24 samples but Hanhyup A line is 13 samples). in total 469 samples were collected from National Institute of Animal Science (Korean Native Chicken (NR, NY, NG, NL and NW), Ogye (NO), Leghorn F,K (NF and NK), Black and Brown cormish (NH and NS), Rhode Island Red C, D (NC and ND), Total is 12 populations] and Hanhyup [H line (HH), F line (HF), G line (HG), V line (HV), S line (HS), W line (HW), Y line (HY), A line (HA), total is 8 populations]. [The allele number were observed 5 (ADL0268) to 20 (MCW0127) each markers. Observed heterozygostiy ($H_{obs}$), expected heterozygosity ($H_{exp}$), polymorphism Information Content (PIC) were observed 0.359 to 0.677, 0.668 to 0.881 and 0.646 to 0.869, respectively. Using these markers, the calculated the heterozygote deficit within chicken line ($F_{is}$) value each population from mean 0.117. Phylogenetic tree showing the genetic relationship among 20 population using standard genetic distance calculated from 27 microsatellite markers. genetic distances revealed the closest (0.175) between NC and ND. on the other hand, Farthest genetic distances (0.710) revealed between NF and HV. STRUCTURE analysis and Principal Components Analysis (PCA) showed that results of similar phylogenetic tree. The expected probability of identity values on random individuals (Total population and only Hanhyup line) was estimated at $8.80{\times}10^{-83}$ and $3.87{\times}10^{-117}$, respectively. In conclusion, This study shows the useful data that be utilized as a basic data of Korean Native Chicken breeding and development for commercial chicken industry to meet the consumer's demand.

Analysis of Genetic Diversity and Population Structure for Core Set of Waxy and Normal Maize Inbred Lines using SSR Markers (SSR 분자마커를 이용한 찰옥수수 및 종실용 옥수수 자식계통들의 핵심집단에 대한 유전적 다양성 및 집단구조 분석)

  • Sa, Kyu Jin;Kim, Jin-Ah;Park, Ki Jin;Park, Jong Yeol;Goh, Byeong Dae;Lee, Ju Kyong
    • Korean Journal of Breeding Science
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    • v.43 no.5
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    • pp.430-441
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    • 2011
  • Maize is divided into two types based on the starch composition of the endosperm in the seed, normal maize(or non-waxy maize) and waxy maize. In this study, genetic diversity and population structure were investigated among 80 waxy maize and normal inbred lines(40 waxy maize inbred lines and 40 normal maize inbred lines) using 50 SSR markers. A total of 242 alleles were identified at all the loci with an average of 4.84 and a range between 2 and 9 alleles per locus. The gene diversity values varied from 0.420 to 0.854 with an average of 0.654. The PIC values varied from 0.332 to 0.838 with an average of 0.602. To evaluate the population structure, STRUCTURE 2.2 program was employed to confirm genetic structure. The 80 waxy and normal maize inbred lines were separated with based on the membership probability threshold 0.8, and divided into groups I, II and admixed group. The 13 waxy maize inbred lines were assigned to group I. The 45 maize inbred lines including 7 waxy maize inbred lines and 38 normal maize inbred lines were assigned to group II. The 22 maize inbred lines with 20 waxy maize inbred lines and 2 normal maize inbred lines were contained in the admixed group. The cluster tree generated using the described SSR markers recognized three major groups at 31.7% genetic similarity. Group I included 40 waxy maize inbred lines and 11 normal maize inbred lines, and Group II included 27 normal maize inbred lines. Group III consist of only 2 normal maize inbred lines. The present study has demonstrated the utility of SSR analysis for the study of genetic diversity and the population structure among waxy and normal maize inbred lines. The information obtained from the present studies would be very useful for designing efficient maize breeding programs in Maize Experiment Station, Kangwon Agricultural Research and Extension Services.

Molecular Analysis and Enzymatic Characterization of Cathepsin B from Olive Flounder (Paralichthys olivaceus) (넙치 카텝신 B의 분자생물학적 분석 및 효소학적 특성 연구)

  • Jo, Hee-Sung;Kim, Na-Young;Lee, Hyung-Ho;Chung, Joon-Ki
    • Journal of Fisheries and Marine Sciences Education
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    • v.26 no.3
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    • pp.543-552
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    • 2014
  • Papain family중 하나인 cysteine protease는 근골격계 질환 치료를 위한target molecule로 인식 되어왔으며 Cathepsin B 는 단백질 분해의 초기과정에 관여하는 cysteine proteases 중 하나이다. 본 연구는 넙치의 cathepsin B 유전자의 발현 양상과 넙치 cathepsin B(PoCtB)의 클로닝, 발현 및 효소특성을 분석하였다. cDNA Library Screening을 이용하여 넙치의 cDNA를 클로닝하였다. 넙치의 동정된 cathepsin B 유전자는 993bp의 open reading frame과 330개의 아미노산으로 이루어져있다. Cathepsin B의 propeptide region 내에 GNFD motif와 occluding loop 가 존재함으로써 이것이 명백하게 cathepsin B group이라는 것을 보여주며, 계통 유전학적 분석 결과 다른 종의 cathepsin B에 비해 초창기에 분화되어 나온 것으로 사료된다. mature enzyme인 maPoCtB은 fusion protein인 glutathione S-transferase를 포함하는 pGEX-4T-1 vector에 삽입하여 E.coli 균주인 $DH5{\alpha}$ 내에 발현시켰다. 재조합 단백질인 PoCtB을 과발현 시킨 결과 53kDa의 분자량을 가진다. 넙치 cathepsin B 활성은 Z-Arg-Arg-AMC와 같은 fluorogenic 펩타이드 기질을 이용하여 측정되었고 적정 pH는 pH.7.5 이다.

Web-based Research Assistant Tools for Analysis of Microbial Diversity (미생물 다양성 분석을 위한 웹 기반의 생물정보도구 개발)

  • 강병철;김현진;박준형;박희경;김철민
    • Proceedings of the Korean Institute of Intelligent Systems Conference
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    • 2004.04a
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    • pp.93-96
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    • 2004
  • 생태학, 환경공학, 임상진단 둥 여러 생물학 분야에서 미생물의 다양성 연구의 중요성이 대두되고 그 연구가 점증하고 있다. 특히 16S rRNA를 분자지표로한 DNA 염기서열 분석방법이 널리 사용되고 있다. 본 논문에서는 16S rRNA의 염기서열 분석과정을 각 단계별로 자동화하고, 생물학자들의 결과 판단이나 사용상의 편의를 도모하기 위하여 웹기반의 미생물 다양성 분석 어플리케이션을 개발하였다. 개발을 위하여 단계별 자동화 및 인터페이스 개발에 적합한 폴더 프로세스-필터 모델을 고안하고 적용하였다. 제공되는 생물정보분석도구는 서열입력, 서열방향교정, 다중서열정렬 및 가시화, 서열동정 등의 분석등이 있으며, 각 결과는 계통분류도구와 호환가능하도록 하였다. 또한 신생아의 장내 세균총에 대한 분석을 수행하여 개발된 도구의 유용성을 확인하였다. 개발된 웹 에플리케이션은 리눅스 시스템 상에서 Perl 과 CGI를 이용하였으며, http: //home.pusan.ac.kr/~genome/tools/rat.htm으로 접속하여 사용할 수 있다.

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