• Title/Summary/Keyword: Banding-methods

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Molecular Analysis of Bacterial Community Structures in Paddy Soils for Environmental Risk Assessment with Two Varieties of Genetically Modified Rice, Iksan 483 and Milyang 204

  • Kim, Min-Cheol;Ahn, Jae-Hyung;Shin, Hye-Chul;Kim, Tae-Sung;Ryu, Tae-Hun;Kim, Dong-Hern;Song, Hong-Gyu;Lee, Geon-Hyoung;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.207-218
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    • 2008
  • The impacts of planted transgenic rice varieties on bacterial communities in paddy soils were monitored using both cultivation and molecular methods. The rice field plot consisted of eighteen subplots planted with two genetically modified (GM) rice and four non-GM rice plants in three replicates. Analysis with denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA genes revealed that the bacterial community structures were quite similar to each other in a given month, suggesting that there were no significant differences in bacterial communities between GM and non-GM rice soils. The bacterial community structures appeared to be generally stable with the seasons, as shown by a slight variation of microbial population levels and DGGE banding patterns over the year. Comparison analysis of 16S rDNA clone libraries constructed from soil bacterial DNA showed that there were no significant differences between GM and non-GM soil libraries but revealed seasonal differences of phyla distribution between August and December. The composition profile of phospholipid fatty acids (PLFA) between GM and non-GM soils also was not significantly different to each other. When soil DNAs were analyzed with PCR by using primers for the bar gene, which was introduced into GM rice, positive DNA bands were found in October and December soils. However, no bar gene sequence was detected in PCR analysis with DNAs extracted from both cultured and uncultured soil bacterial fractions. The result of this study suggested that, in spite of seasonal variations of bacterial communities and persistence of the bar gene, the bacterial communities of the experimental rice field were not significantly affected by cultivation of GM rice varieties.

Genomic DNA Chip: Genome-wide profiling in Cancer

  • 이종호
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2001.10a
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    • pp.61-86
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    • 2001
  • All cancers are caused by abnormalities in DNA sequence. Throughout life, the DNA in human cells is exposed to mutagens and suffers mistakes in replication, resulting in progressive, subtle changes in the DNA sequence in each cell. Since the development of conventional and molecular cytogenetic methods to the analysis of chromosomal aberrations in cancers, more than 1,800 recurring chromosomal breakpoints have been identified. These breakpoints and regions of nonrandom copy number changes typically point to the location of genes involved in cancer initiation and progression. With the introduction of molecular cytogenetic methodologies based on fluorescence in situ hybridization (FISH), namely, comparative genomic hybridization (CGH) and multicolor FISH (m-FISH) in carcinomas become susceptible to analysis. Conventional CGH has been widely applied for the detection of genomic imbalances in tumor cells, and used normal metaphase chromosomes as targets for the mapping of copy number changes. However, this limits the mapping of such imbalances to the resolution limit of metaphase chromosomes (usually 10 to 20 Mb). Efforts to increase this resolution have led to the "new"concept of genomic DNA chip (1 to 2 Mb), whereby the chromosomal target is replaced with cloned DNA immobilized on such as glass slides. The resulting resolution then depends on the size of the immobilized DNA fragments. We have completed the first draft of its Korean Genome Project. The project proceeded by end sequencing inserts from a library of 96,768 bacterial artificial chromosomes (BACs) containing genomic DNA fragments from Korean ethnicity. The sequenced BAC ends were then compared to the Human Genome Project′s publicly available sequence database and aligned according to known cancer gene sequences. These BAC clones were biotinylated by nick translation, hybridized to cytogenetic preparations of metaphase cells, and detected with fluorescein-conjugated avidin. Only locations of unique or low-copy Portions of the clone are identified, because high-copy interspersed repetitive sequences in the probe were suppressed by the addition of unlabelled Cotl DNA. Banding patterns were produced using DAPI. By this means, every BAC fragment has been matched to its appropriate chromosomal location. We have placed 86 (156 BAC clones) cytogenetically defined landmarks to help with the characterization of known cancer genes. Microarray techniques would be applied in CGH by replacement of metaphase chromosome to arrayed BAC confirming in oncogene and tumor suppressor gene: and an array BAC clones from the collection is used to perform a genome-wide scan for segmental aneuploidy by array-CGH. Therefore, the genomic DNA chip (arrayed BAC) will be undoubtedly provide accurate diagnosis of deletions, duplication, insertions and rearrangements of genomic material related to various human phenotypes, including neoplasias. And our tumor markers based on genetic abnormalities of cancer would be identified and contribute to the screening of the stage of cancers and/or hereditary diseases

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Analysis of Archaeal Community in Autotrophic Perchlorate-degrading Enrichment Culture (독립영양 방식으로 퍼클로레이트를 분해하는 농화배양 내 고세균 군집 분석)

  • Kim, Young-Hwa;Do, Sanghyun;So, Hyunseung;Been, Junwon;Sung, Haechan;Ji, Sungchan;Son, Myunghwa;Ahn, Yeonghee
    • Journal of Life Science
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    • v.27 no.4
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    • pp.435-441
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    • 2017
  • Perchlorate ($ClO_4^-$) is an emerging contaminant detected in soil, groundwater, and surface water. Previous study revealed bacterial community in the enrichment culture tdegraded perchlorate using elemental sulfur as an electron donor. Quantitative and qualitative molecular methods were employed in this study to investigate archaeal community in the enrichment culture. Real-time qPCR showed that archaeal 16S rRNA gene copy number in the culture was about 1.5% of bacterial 16S rRNA gene copy number. This suggested that less archaea were adapted to the environment of the enrichment culture and bacteria were dominant. DGGE banding pattern revealed that archaeal community profile of the enrichment culture was different from that of the activated sludge used as an inoculum for the enrichment culture. The most dominant DGGE band of the enrichment culture was affiliated with Methanococci. Further research is necessary to investigate metabolic role of the dominant archaeal population to better understand microbial community in the perchlorate-reducing enrichment culture.

Improvement of PCR Amplification Bias for Community Structure Analysis of Soil Bacteria by Denaturing Gradient Gel Electrophoresis

  • Ahn, Jae-Hyung;Kim, Min-Cheol;Shin, Hye-Chul;Choi, Min-Kyeong;Yoon, Sang-Seek;Kim, Tae-Sung;Song, Hong-Gyu;Lee, Geon-Hyoung;Ka, Jong-Ok
    • Journal of Microbiology and Biotechnology
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    • v.16 no.10
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    • pp.1561-1569
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    • 2006
  • Denaturing gradient gel electrophoresis (DGGE) is one of the most frequently used methods for analysis of soil microbial community structure. Unbiased PCR amplification of target DNA templates is crucial for efficient detection of multiple microbial populations mixed in soil. In this study, DGGE profiles were compared using different pairs of primers targeting different hypervariable regions of thirteen representative soil bacteria and clones. The primer set (1070f-1392r) for the E. coli numbering 1,071-1,391 region could not resolve all the 16S rDNA fragments of the representative bacteria and clones, and moreover, yielded spurious bands in DGGE profiles. For the E. coli numbering 353-514 region, various forward primers were designed to investigate the efficiency of PCR amplification. A degenerate forward primer (F357IW) often yielded multiple bands for a certain single 16S rDNA fragment in DGGE analysis, whereas nondegenerate primers (338f, F338T2, F338I2) differentially amplified each of the fragments in the mixture according to the position and the number of primer-template mismatches. A forward primer (F352T) designed to have one internal mismatch commonly with all the thirteen 16S rDNA fragments efficiently produced and separated all the target DNA bands with similar intensities in the DGGE profiles. This primer set F352T-519r consistently yielded the best DGGE banding profiles when tested with various soil samples. Touchdown PCR intensified the uneven amplification, and lowering the annealing temperature had no significant effect on the DGGE profiles. These results showed that PCR amplification bias could be much improved by properly designing primers for use in fingerprinting soil bacterial communities with the DGGE technique.

Association of polymorphisms in bone morphogenetic protein receptor-1B gene exon-9 with litter size in Dorset, Mongolian, and Small Tail Han ewes

  • Jia, Jianlei;Chen, Qian;Gui, Linsheng;Jin, Jipeng;Li, Yongyuan;Ru, Qiaohong;Hou, Shengzhen
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.7
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    • pp.949-955
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    • 2019
  • Objective: The present study was to investigate the association of polymorphisms in exon-9 of the bone morphogenetic protein receptor-1B (BMPR-1B) gene (C864T) with litter size in 240 Dorset, 232 Mongolian, and 124 Small Tail Han ewes. Methods: Blood samples were collected from 596 ewes and genomic DNA was extracted using the phenol: chloroform extraction method. The 304-bp amplified polymerase chain reaction product was analyzed for polymorphism by single-strand conformation polymorphism method. The genotypic frequency and allele frequency of BMPR-1B gene exon-9 were computed after sequence alignment. The ${\chi}^2$ independence test was used to analyze the association of genotypic frequency and litter size traits with in each ewe breed, where the phenotype was directly treated as category. Results: The results indicated two different banding patterns AA and AB for this fragment, with the most frequent genotype and allele of AA and A. Calculated Chi-square test for BMPR-1B gene exon-9 was found to be more than that of p value at the 5% level of significance, indicating that the population under study was in Hardy-Weinberg equilibrium for all ewes. The ${\chi}^2$ independence test analyses indicated litter size differences between genotypes was not the same for each breed. The 304-bp nucleotide sequence was subjected to BLAST analysis, and the C864T mutation significantly affected litter size in singletons, twins and multiples. The heterozygosity in exon-9 of BMPR-1B gene could increase litter size for all the studied ewes. Conclusion: Consequently, it appears that the polymorphism BMPR-1B gene exon-9 detected in this study may have potential use in marker assisted selection for litter size in Dorset, Mongolian, and Small Tail Han ewes.

The spectrum of 5p deletion in Korean 20 patients with Cri du chat syndrome (한국인 묘성증후군 20명 환자에서의 5p 결실 양상 분석)

  • Park, Sang-Jin;Kim, Sook-Ryung;Baek, Kum-Nyeo;Yoon, Joon-No;Jeong, Eun-Jeong;Kown, Ji-Eun;Kim, Hyon-J.
    • Journal of Genetic Medicine
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    • v.4 no.2
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    • pp.133-141
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    • 2007
  • Purpose : Cri-du-Chat syndrome (CdCs) is a rare but clinically recongnizable condition with an estimated incidence of 1:50,000 live births. The clinical characteristics of the syndrome include severe psychomotor and mental retardation, microcephaly, hypertelorism, hypotonia, and slow growth. Also the size of the chromosome 5p deletion ranges were known from the region 5p13 to the terminal region. In this study, we report the spectrum of 5p deletion in Korean 20 pts. with CdCs and genotype-phenotype associations in CdCs. Methods : In order to delineate genotype-phenotype correlation, molecular cytogenetic studies including GTG banding and clinical characterization were performed on Korean 20 pts with CdCs including parents. CGH array and Fluorescence in situ hybridization (FISH) analysis were used to confirm a terminal deletion karyotype and map more precisely the location of the deletion breakpoint. Results : Molecular analysis of the spectrum of 5p deletion revealed 9 pts (45%) with a del (5)(p14), 7 pts. (35%) a del (5)(p13), 3 pts. (15%) a del (5)(p15.1) and 1 pt. (5%) a del (5)(p15.2) in 20 pts with CdCs. 4(20%)pts were identified to have additional chromosome abnormalites of deficiency and duplication involving chromosomes of 6, 8, 18, & 22. Parental study identified 3 familial case (2 paternal and 1 maternal origin) showing parents being a balanced translocation carrier. And the comparison study of the deletion break points among these 20 pts. with their phenotype has showed the varying clinical pheno-types in the CdCs critical region. Conclusion : The characterization of 5p deletion including parental study may help to delineate the genotypephenotype correlation in CdCs. Also these molecular cytogenetic analyses will be able to offer better information for accurate genetic diagnosis in CdCs and further make possible useful genetic counseling in pts. and family.

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Clinical Characteristics and Genetic Analysis of Prader-Willi Syndrome (Prader-Willi 증후군의 임상 양상 및 유전학적 진단에 관한 고찰)

  • Lee, Ji Eun;Moon, Kwang Bin;Hwang, Jong Hee;Kwon, Eun Kyung;Kim, Sun Hee;Kim, Jong Won;Jin, Dong Kyu
    • Clinical and Experimental Pediatrics
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    • v.45 no.9
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    • pp.1126-1133
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    • 2002
  • Purpose : Prader-Willi syndrome(PWS) is a complex disorder affecting multisystems with characteristic clinical features. Its genetic basis is an expression defect in the paternally derived chromosome 15q11-q13. We analyzed the clinical features and genetic basis of PWS patients for early detection and treatment. Methods : We retrospectively studied 24 patients with PWS in Department of Pediatrics, Samsung Medical Center, from September 1997 to September 2001. We performed cytogenetic and molecular genetic techniques using high resolution GTG banding techniques, fluorescent in situ hybridization and methylation-specific PCR for CpG island of SNRPN gene region. Results : The average birth weight of PWS patients was $2.67{\pm}0.47kg$ and median age at diagnosis was 1.3 years. The average height and weight of PWS patients under one year at diagnostic time were located in a 3-10 percentile relatively, and a rapid weight gain was seen between two and six years. Feeding problems in infancy and neonatal hypotonia were the two most consistently positive major criteria in over 95% of the patients. In 18 of the 24 cases(75%), deletion of chromosome 15q11-q13 was demonstrated and one case among 18 had an unbalanced 14;15 translocation. In four cases without any cytogenetic abnormality, it may be considered as maternal uniparental disomy and the rest showed another findings. Conclusion : We suggest diagnostic testing for PWS in all infants/neonates with unexplained feeding problems and hypotonia. It is necessary for clinically suspicious patients to undergo an early genetic test. As the genetic basis of PWS was heterogenous and complex, further study is required.

Effect of Timing and Placement of N Fertilizer Application for Increased Use Efficiency - Principle and Practice (열대지역(熱帶地域)에 있어서 질소비료(窒素肥料)의 시용시기(施用時期)와 시비위치(施肥位置)가 비료효율(肥料效率)에 미치는 영향(影響) - 원리(原理)와 실제(實際))

  • Hong, Chong-Woon
    • Korean Journal of Soil Science and Fertilizer
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    • v.20 no.3
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    • pp.285-299
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    • 1987
  • Timing and placement of fertilizer applications are two managerial means to improve the fertilizer use efficiency. The relative importance of these two means is determined by the application rate. With the realistic rate of N application recommended to the small farmers in the tropics, at present and in the near future, basal application in right manner, seems to be more important than split application at different times. In wetland rice soils, deep placement by whatever available means is desirable. But in the situations where perfect deep placement is very difficult to implement, the whole-layer application may be worth trying, until better methods become available. In rainfed uplands, N fertilizer application plans should be contingent upon the amount and distribution of rainfall: apply a less risky rate as subsurface banding near the crop rows to start with; then, depending upon the rainfall prospects in the season, apply or omit the additional dose. Because the patterns of crop response to N fertilizer can be significantly different between the research farms and farmers' fields, it seems imperative to have information on the patterns of crop response to N under farmers' management conditions, for the development of realistic fertilizer application recommendations. To enable the farmers to adopt improved fertilizer application technologies, it is essential to develop and make available to farmers convenient fertilizer applicators. Past experience with the improved fertilizer use technologies indicates that, in the long run, the development of fertilizers that are not only effective and convenient for farmers to use but also easy to produce without major modifications of existing fertilizer production systems is the ultimate solution to the problem of low N fertilizer use efficiency.

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지점우량 자료의 분포형 설정과 내용안전년수에 따르는 확률강우량에 관한 고찰 - 국내 3개지점 서울, 부산 및 대구를 중심으로 -

  • Lee, Won-Hwan;Lee, Gil-Chun;Jeong, Yeon-Gyu
    • Water for future
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    • v.5 no.1
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    • pp.27-36
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    • 1972
  • This thesis is the study of the rainfall probability depth in the major areas of Korea, such as Seoul, Pusan and Taegu. The purpose of the paper is to analyze the rainfall in connection with the safe planning of the hydraulic structures and with the project life. The methodology used in this paper is the statistical treatment of the rainfall data in the above three areas. The scheme of the paper is the following. 1. The complementation of the rainfall data We tried to select the maximm values among the values gained by the three methods: Fourier Series Method, Trend Diagram Method and Mean Value Method. By the selection of the maximum values we tried to complement the rainfall data lacking in order to prevent calamities. 2. The statistical treatment of the data The data are ordered by the small numbers, transformed into log, $\sqrt{}, \sqrt[3]{}, \sqrt[4], and$\sqrt[5], and calculated their statistical values through the electronic computer. 3. The examination of the distribution types and the determination of the optimum distibution types By the $x^2-Test$ the distribution types of rainfall data are examined, and rejected some part of the data in order to seek the normal rainfall distribution types. In this way, the optimum distribution types are determined. 4. The computation of rainfall probability depth in the safety project life We tried to study the interrelation between the return period and the safety project life, and to present the rainfall probability depth of the safety project life. In conclusion we set up the optimum distribution types of the rainfall depths, formulated the optimum distributions, and presented the chart of the rainfall probability depth about the factor of safety and the project life.ct life.

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Weight loss effects of Bariatric Surgery after nutrition education in extremely obese patients (고도비만환자에서 베리아트릭 수술 (Bariatric Surgery) 후 영양교육이 체중감량에 미치는 효과)

  • Jeong, Eun-Ha;Lee, Hong-Chan;Yim, Jung-Eun
    • Journal of Nutrition and Health
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    • v.48 no.1
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    • pp.30-45
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    • 2015
  • Purpose: This study was planned to determine the characteristics of extremely obese patients during Bariatric surgery and to evaluate how the difference in the number of postsurgical personal nutritional educations they received affected the weight loss. Methods: This is a retrospective study on the basis of the medical records of extremely obese patients for 15 months after receiving gastric banding. A total of 60 people were selected as the study subjects and they were divided into the Less Educated Group and the More Educated Group according to the average number of personal nutritional educations they received. We investigated both groups to determine the general characteristic, health related lifestyle habits, obesity related complications and symptoms in possession, and eating habits before their surgery, the body composition measurement result, obesity determination indices at 1, 3, 6, 9, 12, and 15 months before and after their surgery, and the biochemical parameters at 6 months before and after their surgery. Results: Body fat and weight showed rapid reduction until 6 months after the surgery, but thereafter reduced slowly depending on the result of body composition measurement. Regarding body fat and weight, the More Educated Group, who received nutrition education more often, showed significantly lower levels than the Less Educated Group at 15 months after surgery. Regarding BMI and degree of obesity, the More Educated Group showed significantly lower levels than the Less Educated Group at 15 months after surgery. Here, we were assured that BMI is reversely proportional to the number of personal nutritional educations at 15 months, which is more outstanding after surgery than before surgery. Conclusion: Long-term nutritional education is a key factor for the extremely obese patient in maintaining the effects of Bariatric surgery on weight and body fat reduction onwards. In the next stage, considering the characteristics of the study subjects, adoption of individual nutrition education is recommended for postsurgical prospective arbitration of obesity in order to monitor blood pressure, obesity related complications, symptoms in possession, and how eating habits and health related life habits change, and to judge the actual effect of the nutritional education method at the same time.