• Title/Summary/Keyword: Copy Numbers

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Progeny Analysis and Selection of Tomato Transformants with patII Gene linked to Inherent Disease Resistance Gene (제초제 저항성 유전자와 기존 병 저항성 유전자가 연관된 형질전환 토마토 개체 선발 및 후대분석)

  • Ahn, Soon-Young;Kang, Kwon-Kyoo;Yun, Hae-Keun;Park, Hyo-Guen
    • Horticultural Science & Technology
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    • v.29 no.4
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    • pp.345-351
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    • 2011
  • This study was carried out to develop a model system using selection method for disease resistant plant breeding programs using a herbicide bialaphos-resistant patII gene as a gene-based marker. Spraying bialaphos could eliminate the susceptible plants from the segregating populations such as ${F_2}^{\prime}s$ and thereafter. Tomato cv. Momotaro-yoke was transformed with patII gene 60 independent transformants were acquired. Total 42 transformants were analyzed in transgene copy numbers by Southern blotting and the segregation ratios for the bialaphos resistance. Statistical analysis revealed that the transgene copy numbers and the segregation ratios were not always coincided, especially having the tendency of underestimating the real numbers of the transgenes in the multicopy lines. A two-stepwise screening method was applied to select $T_1$ tomato plants which linked the transgenic patII to a disease resistance gene (I2 and Ve). Based on the resistant to susceptible ratios, T-20 plant was finally selected due to the estimated linkage 12-13 cM between the patII gene to the I2 gene on chromosome 11. This newly developed system could be applied to any economical crop in breeding programs.

A Study of Eye Blinking : I (눈의 깜박임에 대한 연구 : I)

  • Gong, Myeong-Ok;Kim, Yeon-Min
    • Journal of the Ergonomics Society of Korea
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    • v.4 no.1
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    • pp.7-10
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    • 1985
  • Eye blinking in case of three visual tasks is studied; wall hard-copy and color monitor. The eye blinking is checked by eye-status detector. The data observed are (1) the average number of eye blinking (2) the average time of eye blinking for three minutes in case of all three visual tasks. The results are as follows: (1) The average times (or numbers) of eye blinking are different among three tasks. (2) The average time (or number) of eye blinking of wall is different from hard-copy and color monitor tasks. (3) But the average time (or number) of eye blinking of hard-copy is not different from that of color monitor.

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Distribution of Strain Types and Function of P Transposable Element in Natural Populations of Drosophila melanogaster (초파리 자연집단의 P 전이인자에 대한 계통형 분포와 기능에 관한 연구)

  • 김지식;권도형추종길
    • The Korean Journal of Zoology
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    • v.38 no.2
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    • pp.177-185
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    • 1995
  • 두 지역에서 채집한 초파리 자연집단에 대하여 난소발생이상 실험에 의한 P 인자 활성과 세포질형을 분석하여 P 전이인자의 계통형을 조사하였다 전체 238 isofemale line을 조사한 결과 strong P와 true M은 존재하지 않았고, 0(weak P)와 M'(pseudo M) strain이 전체의 98.74%를 차지하여 가장 우세하게 분포하고 있었다. P$\pi$25.1 probe를 이용한 in situ hybridization을 행하여 P 전이인자의 copy수를 조사한 결과 평균 42.12개로 나타났으며, 0와 M'의 계통형 간에 유의적인 차이는 없었다 그러나 염색체 firm당 COPy수는 X염색체가 상염색체의 좌 우 각 arm보다 다소 높게 분포하고 있었고. 염색체상 P 전이인자의 삽입부위에 대한 특이적 좌위는 존재하지 않았다 P 전이인자의 분자구조에 대한 변이형을 조사하기 위하여 southern blot hybridization을 행한 결과 2.9kb의 완전한 크기의 분자를 포함하여 여러종류의 단편들이 확인되었다 조사한 모든 isofemale line에서 KP(1. 15kb)인자를 포함하고 있었으며 이들 KP인자가 P-M System의 난소발생이상을 표현하는데 있어 억제적 작용을 하는 것으로 판단되었다.

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재조합 효모를 이용한 항혈전 단백질 히루딘 발효 생산공정의 최적화

  • Kim, Myeong-Dong;Gang, Hyeon-A;Lee, Sang-Gi;Seo, Jin-Ho
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.99-102
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    • 2001
  • Recombinant Saccharomyces cerevisiae strains harboring various copy numbers of hirudin gene were developed to study dependency of hirudin expression level on its gene copy number. A linear relationship between the copy number of hirudin expression cassette and hirudin expression level was observed up to 10 copies. A double <5-integration vector truncated wi 디 1 the unnecessary bacterial genes before yeast transformation showed a four-fold increase in transformation efficiency and a 1.3-fold enhancement in hirudin expression level compared with a single <5 system. Gratuitous hirudin expression strain was developed by disrupting the GALl gene of S. cerevisiae. Glucose that was fed in a limited manner effectively supported cell growth and hi겨din expression by the gratuitous strain. Effects of methanol concentrations on hirudin production in recombinant Hansenula polymorpha were investigated in continuous and fed-batch cultures. At a steady-state of continuous culture, an optimum methanol concentration of 1.7 g/L was determined at a dilution rate of 0.18 $h^{-1}$ with 1.8 mg/L ${\cdot}$ h hirudin productivity.

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A SPECTROSCOPIC STUDY OF THE OPTICAL PROPERTIES OF VARIOUS PAPERS MADE FROM RECLAIMED FIBRES

  • Ingegerd Forsskahl;Carola Olkkonen;Henrik Tylli
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 1999.04b
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    • pp.296-301
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    • 1999
  • Different papers such as toilet and towel tissue, newsprint and copy paper made from reclaimed fibres were characterized using UV-VIS reflectance and fluorescence spectroscopy. The emission properties of the extracts of the papers were also evaluated. Photochemical irradiation at ambient temperatures, and thermal aging of the copy papers at 105$^{\circ}C$ and 150$^{\circ}C$ were performed. Post-colour numbers (PC) were calculated from the reflectance values measured from the corresponding UV-VIS reflectance spectra. Light absorption coefficient spectra were calculated before and after irradiation, and the changes in absorption coefficient were related to the content of chromophores in the papers. Photochemical and thermal discolouration of the copy papers, similar to that of virgin fibres, was readily observed. The influence of changes in the concentrations of fluorescent brightening agents and dyes on the spectral properties was also assessed. Photostabilization studies of the copy papers were performed with the use of polymer additives polyethylene glycol (PEG) and polytetrahydrofuran (PTHF).

Comparison of Methods for Detecting and Quantifying Variation in Copy Numbers of Duplicated Genes

  • Jeon, Jin-Tae;Ahn, Sung-Jin
    • Communications for Statistical Applications and Methods
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    • v.16 no.6
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    • pp.1037-1046
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    • 2009
  • Copy number variations(CNVs) are known as one of the most important factors in susceptibility to genetic disorders because they affect expression levels of genes. In previous studies, pyrosequencing, mini-sequencing real-time polymerase chain reaction(PCR), invader assays and other techniques have been used to detect CNVs. However, the higher the copy number in a genome, the more difficult it is to resolve the copies, so a more accurate method for measuring CNVs and assigning genotype is needed. PCR followed by a quantitative oligonucleotide ligation assay(qOLA) was developed for quantifying CNVs. The aim of this study was to compare the two methods for detecting and quantifying the CNVs of duplicated gene: the published pyrosequencing assay(pyro_CNV) and the newly developed qOLA_CNV. The accuracy and precision of the assay were evaluated for porcine KIT, which was selected as a model locus. Overall, the root mean squares(RMSs) of bias and standard deviation of qOLA_CNV were 2.09 and 0.45, respectively. These values are less than half of those of pyro CNV.

The change of PHP index and copy numbers of periodontal bacteria in patients with professional periodontal prevention management (PHP INDEX에 따른 전문가 치주예방관리 환자의 치주질환 원인균 copy 수 변화)

  • Mi-Young, Jin;Byeng-Chul, Yu
    • Journal of Korean Dental Hygiene Science
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    • v.5 no.2
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    • pp.155-164
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    • 2022
  • Background: By regulating the factors that contribute to oral diseases in a healthy way, oral health can be maintained and prevented. Methods: The general characteristics, PHP index, oral health behavior, and clinical periodontal index of each group were calculated by frequency analysis, and a cross-analysis (χ2 test) was conducted to assess the homogeneity of the general characteristics, oral health behavior, and clinical periodontal index of the study subjects. An Oral bacteriological examination was performed by gargling with saliva. Results: The expert periodontal prevention group showed a decrease in the copy number of periodontal disease causative bacteria, and A. actinomycetemcomitans, P. gingivalis, T. forsythus, andT. denticolashowed a significant difference before and after treatment (p=0.021). In the periodontal treatment group, A. actinomycetemcomitans, P. gingivalis, T. forsythus, andT. denticolaall showed a decrease in copy number, but there was no significant difference. Conclusions: This study showed professional periodontal prevention management had some effect on periodontal bacterial reduction.

Gene Duplications Revealed during the Process of SNP Discovery in Soybean[Glycine max(L.) Merr.]

  • Cai, Chun Mei;Van, Kyu-Jung;Lee, Suk-Ha
    • Journal of Crop Science and Biotechnology
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    • v.10 no.4
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    • pp.237-242
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    • 2007
  • Genome duplication(i.e. polyploidy) is a common phenomenon in the evolution of plants. The objective of this study was to achieve a comprehensive understanding of genome duplication for SNP discovery by Thymine/Adenine(TA) cloning for confirmation. Primer pairs were designed from 793 EST contigs expressed in the roots of a supernodulating soybean mutant and screened between 'Pureunkong' and 'Jinpumkong 2' by direct sequencing. Almost 27% of the primer sets were failed to obtain sequence data due to multiple bands on agarose gel or poor quality sequence data from a single band. TA cloning was able to identify duplicate genes and the paralogous sequences were coincident with the nonspecific peaks in direct sequencing. Our study confirmed that heterogeneous products by the co-amplification of a gene family member were the main cause of obtaining multiple bands or poor quality sequence data in direct sequencing. Counts of amplified bands on agarose gel and peaks of sequencing trace suggested that almost 27% of nonrepetitive soybean sequences were present in as many as four copies with an average of 2.33 duplications per segment. Copy numbers would be underestimated because of the presence of long intron between primer binding sites or mutation on priming site. Also, the copy numbers were not accurately estimated due to deletion or tandem duplication in the entire soybean genome.

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Effects of feed intake on the diversity and population density of homoacetogens in the large intestine of pigs

  • Matsui, Hiroki;Mimura, Ayumi;Maekawa, Sakiko;Ban-Tokuda, Tomomi
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.12
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    • pp.1907-1913
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    • 2019
  • Objective: Homoacetogens play important roles in the production of acetate in the large intestine of monogastric mammals. However, their diversity in the porcine large intestine is still unknown. Marker gene analysis was performed to assess the effects of energy level on the diversity and population densities of homoacetogens in porcine feces. Methods: Crossbred pigs were fed high or low energy-level diets. The high-intake (HI) diet was sufficient to allow a daily gain of 1.2 kg. The low-intake (LI) diet provided 0.6 times the amount of energy as the HI diet. Genetic diversity was analyzed using formyltetrahydrofolate synthetase gene (FHS) clone libraries derived from fecal DNA samples. FHS DNA copy numbers were quantified using real-time polymerase chain reaction. Results: A wide variety of FHS sequences was recovered from animals in both treatments. No differences in FHS clone libraries between the HI and LI groups were found. During the experimental period, no significant differences in the proportion of FHS copy numbers were observed between the two treatment groups. Conclusion: This is the first reported molecular diversity analysis using specific homoacetogen marker genes from the large intestines of pigs. There was no observable effect of feed intake on acetogen diversity.

Flanking Sequence and Copy-Number Analysis of Transformation Events by Integrating Next-Generation Sequencing Technology with Southern Blot Hybridization

  • Qin, Yang;Woo, Hee-Jong;Shin, Kong-Sik;Lim, Myung-Ho;Cho, Hyun-Suk;Lee, Seong-Kon
    • Plant Breeding and Biotechnology
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    • v.5 no.4
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    • pp.269-281
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    • 2017
  • With the continual development of genetically modified (GM) crops, it has become necessary to develop detailed and effective molecular characterization methods to select candidate events from a large pool of transformation events. Relative to traditional molecular analysis methods such as the polymerase chain reaction (PCR) and Southern blot hybridization, next generation sequencing (NGS) technology for whole-genome sequencing of complex crop genomes had proven comparatively useful for in-depth molecular characterization. In this study, four transformation events, including one in Bacillus thuringiensis (Bt)-resistant rice, one in resveratrol-producing rice, and two in beta-carotene-enhanced soybeans, were selected for molecular characterization. To merge NGS analysis and Southern blot-hybridization results, we confirmed the transgene insertion sites, insertion construction, and insertion numbers of these four transformation events. In addition, the read-coverage depth assessed by NGS analysis for inserted genes might provide consistent results in terms of inserted T-DNA numbers in case of complex insertion structures and highly duplicated donor genomes; however, PCR-based methods can produce incorrect conclusions. Our combined method provides an effective and complete analytical approach for whole-genome visual inspection of transformation events that require biosafety assessment.