• Title/Summary/Keyword: 분자표지

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Replication Inhibition and Its Recovery/Process in Chinese Hamster Ovary Cells Treated with Methyl Methanesulfonate (Chinese Hamster Ovary세포에 있어 methyl methanesulfonate에 의한 DNA 복제억제와 이의 회복경로)

  • 이천복;이형호;박상대;이치건
    • Environmental Mutagens and Carcinogens
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    • v.9 no.1
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    • pp.33-46
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    • 1989
  • 본 연구는 알킬화제를 처리한 CHO-K1 세포에서 DNA 복제억제와 그 회복과정의 분자론적 기작을 규명할 목적으로 방사선 이중 표지에 의한 DNA 합성율의 측정, 알칼리 자당 농도구배 초원심분리법에 의한 DNA 분자량과 후복제 회복율을 측정하여 다음과 같은 결과를 얻었다. (1) 1mM methyl methanesulfonate (MMS)와 1nM N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) 이하의 낮은 농도의 처리군에서는 DNA 합성율이 급격히 감소하였으나, 2 mM MMS, 2mM MNNG이상의 농도에서는 그 감소양상이 둔화되었다, (2) DNA 합성율은 알킬화제의 처리 직후 감소하였다가 시간경과에 따라 회복되어 처리후 4시간 째에는 대조군 수준 또는 그 이상으로 회복되었다.

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Effects of Testosterone on Adipose Tissue Metabolism (지방조직대사에 대한 testosterone의 영향)

  • Jeong, Sunhyo
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.17 no.12
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    • pp.2995-3000
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    • 2013
  • We investigated the effects of testosterone on the improvement of white adipose tissue explant and its molecular mechanism in adipose tissue of high fat diet-fed male castrated (CAST) mice. The CAST mice treated with testosterone had lower adipose tissue weights, the average size of adipocytes and mRNA levels of $C/EBP{\alpha}$ as well as adipocyte marker genes than the vehicle-treated CAST mice. These results suggest that testosterone prevent the expression of $C/EBP{\alpha}$ and $C/EBP{\alpha}$-mediated adipocyte marker genes, resulting in decreased adipose tissue mass and adipocyte metabolism in male CAST mice. Moreover, this study give a valuable molecular and biological knowledge on testosterone therapy in obese hypogonadal men.

A Study of Influences of p53 Mutation, Cyclin D1 Over Expression, Ki67 Index, Mitotic Index on the Prognosis of Esophageal Squamous Cell Carcinoma (p53 변이, Cyclin D1의 과발현, Ki67 지수, 세포분열지수가 식도의 편평상피암의 예후에 미치는 영향)

  • Lee Hae Won;Cho Suk Ki;Sung Sook Whan;Lee Hyun Joo;Kim Young Tae;Kang Moon Chul;Kim Joo Hyun
    • Journal of Chest Surgery
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    • v.38 no.12 s.257
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    • pp.835-843
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    • 2005
  • Background: Cancer of the esophagus is one of the most malignant tumors with poor prognosis. The p53 gene alteration, over expression of Cyclin D1, and Ki67 index were thought to be the prognostic factors. However, their clinical significances in esophageal squamous cell carcinoma are controversial and p53 accumulation may not correlate with genetic mutation. The current study investigates their prognostic significance in squamous cell carcinoma of the esophagus. Material and Method: The Subjects studied were 124 esophageal squamous cell carcinoma patients who underwent esophagectomy. The mutation of p53, over expression of Cyclin D1, Ki67 labelling index, mitotic index were examined by using an immunohistochemical staining. We compared the results and investigated the correlation with the mutation of p53, overexpression of Cyclin D1, Ki67 labelling index, mitotic index and tumor size, and duration of survival. Result: There was no correlation between the results in immunohistochemical staining according to age, sex, tumor size, Iymph node status, and clinical stage of the disease. Mutant p53 protein was found in 69 cases (55.6$\%$). Median survival time was 21 months in cases with negative for mutant p53 protein and 22 months in positive cases. There was no significant difference in survival (p=0.46). Median survival time was 22 months in cases with negative for Cyclin D1 and 16 months in positive cases (p=0.18). Median and mean survival time was 22 months and 36 months when Ki67 labeling index was 40 or less (102 cases). Median and mean survival was 16 months and 23 months, when Ki67 labeling index was more than 40 (22 cases). There was significant difference in survival rate (p=0.011). Conciusion: Positivity of p53 and cyclin D1 was not useful in predicting the prognosis in our study. There was no significant correlation among mutant p53 protein accumulation, Cyclin D1 over expression, and Ki67 labeling index. However, in several studies, PCR single strand conformational polymorphism analysis of p53 showed a correlation to the prognosis. We thought that there was a significant discordance between p53 gene mutation and mutant p53 protein accumulation. When Ki67 labeling index was more than 40, prognosis was poorer, Ki67 seems to be a prognostic factor in our study. Therefore, we confirmed the possibility of using molecular markers as prognostic factors.

Improvement of Seedling Establishment in Wet Direct Seeding of Rice using the Anaerobic Germination Tolerance Gene Derived from Weedy Photoblastic Rice (잡초벼 PBR 혐기발아 내성 유전자 활용 벼 담수직파 초기 입모 개선)

  • Jeong, Jong-Min;Mo, Youngjun;Baek, Man-Kee;Kim, Woo-Jae;Cho, Young-Chan;Ha, Su-Kyung;Kim, Jinhee;Jeung, Ji-Ung;Kim, Suk-Man
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.3
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    • pp.161-171
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    • 2020
  • Direct seeding is one of the rice seedling establishment methods that is increasingly being practiced by farmers to save labor and reduce costs. However, this method often causes poor germination under flooding conditions after sowing. In this study, we developed japonica elite lines with quantitative trait loci (QTL) associated with anaerobic germination (AG) tolerance to overcome poor germination and seedling establishment in wet direct seeding. The QTL introgression lines were developed from a cross between weedy photoblastic rice as the AG donor and the Nampyeong variety via phenotypic and genotypic selection. Compared to Nampyeong, the survival rates of the selected lines were improved by approximately 50% and 240% under field and greenhouse conditions, respectively. To improve selection efficiency by marker assisted selection, the QTL markers associated with AG tolerance were converted to cleaved amplified polymorphic sequence markers designed based on next-generation sequence analysis. These lines retained similar agronomic traits and yield potential to the parent, Nampyeong. Among these lines, we selected the most promising line, which exhibited high survival rate and good agricultural traits under flooding conditions and named the line as Jeonju643. This line will contribute to breeding programs aiming to develop rice cultivars adapted to wet direct seeding. This study demonstrates the successful application of marker-assisted selection to targeted introgression of anaerobic genes into a premium quality japonica rice variety.

Development of SNP Molecular Markers Related to Seed-hair Characteristic Based on EST Sequences in Carrot (당근 EST 염기서열을 이용한 종자모 형질 관련 SNP 분자표지 개발)

  • Oh, Gyu-Dong;Shim, Eun-Jo;Jun, Sang-Jin;Park, Young-Doo
    • Horticultural Science & Technology
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    • v.31 no.1
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    • pp.80-88
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    • 2013
  • Carrot (Daucus carota L. var. sativa) is one of the most extensively used vegetable crops in the world and a significant source of nutrient because of its high content of ${\beta}$-carotene, well known as the precursor of vitamin A carotenoid. However, seed-hairs generated and elongated from the epidermal cell of seeds inhibit absorption and germination by various factors such as carotol and so on. Accordingly, mechanical hair removal process is essential before commercialization of carrot seeds. Because of this process, producers will have additional losses such as time consuming, manpower, capital and so on. Furthermore, physical damage of seeds causes irregular germination rate. To overcome such cumbersome weaknesses, new breeding program for developing hairless-seed carrot cultivar has been needed and studies for molecular markers related to seed-hair characteristic is needed for a new breeding program. Therefore, in this study, cDNA libraries from seeds of short-hair seed phenotype CT-SMR 616 OP 659-1 line, hairy-seed phenotype CT-SMR 616 OP 677-14 line and short-hair seed phenotype CT-ATR 615 OP 666-13 line, hairy-seed phenotype CT-ATR 615 OP 671-9 were constructed, respectively. Furthermore, 1,248 ESTs in each line, total 4,992 ESTs were sequenced. As a result, 19 SNP sites and 14 SNP sites in each of 2 combinations were confirmed by analyzing these EST sequences from short-hair and hairy-seed lines. Then we designed SNP primer sets from EST sequences of SNP sites for high resolution melting (HRM) analysis. Designed HRM primers were analyzed using hairy seed phenotype CT-SMR 616 OP 1040 line and short-hair seed phenotype CT-SMR 616 OP 1024, 1025, 1026 lines. One set of HRM primers showed specific difference between the melting curves of hairy and short-hair seed phenotype lines. Based on this result, allele-specific (AS) PCR primers were designed for easier selection between hairy-seed carrot and hairless seed carrot. These results of HRM and AS-PCR are expected to be useful in breeding of hairless seed carrot cultivar as a molecular marker.

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.

Wolbachia-mediated Reproductive Alterations in Arthropod Hosts and its use for Biocontrol Program (볼바키아 세균에 의한 절지동물 기주의 생식적 변화와 생물적방제 프로그램에 이용 방안)

  • Rostami, Elahe;Madadi, Hossein;Abbasipour, Habib;Sivaramakrishnan, Shiva
    • Korean journal of applied entomology
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    • v.55 no.2
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    • pp.177-188
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    • 2016
  • The alpha-proteobacterium Wolbachia is one of the most important intracellular symbionts of arthropods. This Gram-negative bacterium is involved in many biological processes and is currently considered as a potential tool for biological control. Wolbachia is a cytoplasmic bacterium, maternally transferred through generations, and to facilitate its success, it has evolved several strategies that manipulate its host reproductive system to increase the number of infected individuals in the host population. The variety of Wolbachia was first recognized using genes wsp, 16S rRNA, ftsZ, gltA and groEL as molecular markers while strain genotypes of Wolbachia are determined of Multilocus sequence typing (MLST) and sequence of amino acid in region, hyper variable regions (HVRs) in protein WSP. Possible uses of the bacteria and their predominant phenotypes in control programs for agricultural pests and human disease vectors have been considered. Phenotypes are known to induce cytoplasmic incompatibility (CI), parthenogenesis induction (PI), feminization (F) and male killing (MK). Finally, applications of the bacterium in control programs of agricultural and medical insect pests have been discussed.

Pickprimer: A Graphic User Interface Program for Primer Design on the Gene Target Region (픽프라이머 : 유전자 목표 구간 탐색 모듈을 포함한 프라이머 제작 그래픽 프로그램)

  • Chung, Hee;Mun, Jeong-Hwan;Lee, Seung-Chan;Yu, Hee-Ju
    • Horticultural Science & Technology
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    • v.29 no.5
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    • pp.461-466
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    • 2011
  • In genetic and molecular breeding studies of plants, researchers need to design various kinds of primers based on their research purposes. So far many kinds of web- or script-based non-commercial programs for primer design are available. Because most of them do not include user interface for multipurpose usage including gene structure prediction and direct target selection on sequences, it has been a laborious work to design primers targeting on the exon or intron regions of interesting genes. Here we report a primer designing graphic user interface program, Pickprimer, that includes gene structure prediction and primer design modules by combining source codes of the Spidey and Primer3 programs. This program provides simple graphic user interface to input sequences and design primers. Genomic sequence and mRNA or coding sequence of genes can be copy and pasted or input as fasta or text files. Based on alignment of the input sequences using the Spidey module, a putative gene structure is graphically visualized along with exon-intron sequences of color codes. Primer design can be easily performed by dragging mouse on the displayed sequences or input primer targeting position with desirable values of primers. The output of designed primers with detailed information is provided by the Primer3 module. PCR evaluation of 24 selected primer sets successfully amplified single amplicons from six Brassica rapa cultivars. The Pickprimer will be a convenient tool for genetic and molecular breeding studies of plants.

Characteristics and Improving Breed of Economic Traits of Korea Native Chicken (한국 재래 닭 품종 특성 및 초기성장 개량을 위한 분자표지 개발)

  • Oh J. D.;Park M. H.;Kong H. S.;Lee H. K.;Jeon G. J.;Yeon S. H.;Sang B. D.;Choi C. H.;Cho B. W.
    • Korean Journal of Poultry Science
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    • v.32 no.1
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    • pp.29-34
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    • 2005
  • This study was conducted to estimate the effects of genotype for chicken major histocompatibility complex (MHC) B-LB genes on economic traits. To detect polymorphism, 400 bp fragments of MHC B-LB genes were obtained and sequenced. After digestions using restriction enzyme Hea III, two restriction enzyme sites were observed. There were two mutations at position 427 and 651 those were decided as Type I and Type II, respectively. Using RFLP analyses, type I were genotyped to TT, TC and CC, and type II to MM, Mm and mm. The relatively higher TC genotype frequencies (0.8) of Type I and Mm genotype frequencies (0.88) of Type II were observed in Korean native chickens. The effects of the genotype on 150 days body weight trait were investigated by the associations of CC and Mm genotypes (P<0.05) in Korean native chickens. This result suggests that a significant association exists between the SNP and 150 days body weight.