• Title/Summary/Keyword: 유전자 삽입

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Quantitative Analysis of Genetically Modified Soybean in Processed Foods Using Real-time PCR (실시간중합효소연쇄반응을 이용한 유전자변형 콩 가공식품의 정량분석)

  • Min, Dong-Myung;Kim, Myo-Young;Jung, Soon-Il;Heo, Mun-Seok;Kim, Jin-Kuk;Kim, Hae-Yeong
    • Korean Journal of Food Science and Technology
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    • v.36 no.5
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    • pp.723-727
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    • 2004
  • Qualitative and quantitative PCR methods were performed to examine detection and quantitation of epsps inserted into genetically modified soybean (GMS) in processed foods, soy milk, tofu, and biji (soybean curd residue). Using PCR amplification to produce two (121 and 330 bp) epsps in GMS, detection limits of GMS in soy milk, tofu, and biji containing 0.01% GMS were measured. For quantitative detection, test samples containing 1, 3, and 5% GMS were measured by real-time PCR method. Results show real-time PCR method is applicable to detect GMS quantitatively in processed foods.

Studies on Mammalian Homolog and Flanking Sequence of Mouse MT Transposon-like Element, Clone MTi7(MTi7) (생쥐의 MT Transposon-like Element, Clone MTi7(MTi7) 유전자의 포유류 Homolog 및 Flanking Sequence에 대한 연구)

  • Kim Young-Hoon;Ko Minsu;Woo Dae-Gyun;Choi Donchan;Lee Kyung-Ah
    • Development and Reproduction
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    • v.7 no.2
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    • pp.119-126
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    • 2003
  • In the previous study, we obtained list of differentially expressed genes between postnatal day 1 and day 5 mouse ovaries using suppression subtractive hybridization(SSH) and found that MT Oansposon-like element, clone MTi7(MTi7) was one of the highly expressed genes in the day 5 mouse ovary(Park et at., 2002). Results of in situ hybridization and RNA interference revealed that the expression of MTi7 is oocyte-specific in the ovary and may be involved in the regulation of oocyte maturation(Park et at., 2003). At present, MTi7 sequence has been known only in the mouse. Therefore, the present study was accomplished 1) to identify MTi7 sequence in the other mammalian species, such as bovine, porcine, rat, and human, and 2) to evaluate the flanking sequence of the mouse MTi7 since it has transposon characteristics. Using ovarian cDNAs derived from low different species, we cloned and identified new MTi7 sequence showing a high degree of sequence homology with the mouse MTi7(87∼98%). By using inverse PCR, we found that the mouse MTi7 may intercalated the beta-carotene 15, 15'-monooxygenase(Bcdo) gene and/or serine protease inhibitor, Kunitz Dpe I(Spint 1) gene. By finding the MTi7 sequences in the other mammalian species and the flanking gene of the MTi7 in mouse, it is expected to reveal the role(s) of MTi7 in the oogenesis as well as folliculogenesis in the near future.

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Expression of the lux Genes in Escherichia coli for the Basis of Development of Biosensor (바이오센서 개발을 위한 발광 유전자의 대장균에서의 발현 시스템 분석)

  • Cho, Mi-Mi;Kim, Young-Doo;Kang, Kyung-Sook;Kim, Sook-Kyung;Yang, In-Chul;Park, Sang-Ryoul;Lee, Chan-Yong
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.419-424
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    • 2009
  • To provide the basis of biosensor based on the lux genes from bioluminescent bacteria of Photobacterium leiognathi and Vibrio harveyi, we test the expression of lux genes in several strains of Escherichia coli. The expression of the recombinant plasmid of PlXba.pT7-3, containing all lux genes requiring for light emission without adding substrate, in E. coli 43R was so strong to see the blue-green light in single colony as well as in the alginate immobilized cell. In addition, the light intensity was decreased by adding heavy metal ion such as cadmium and zinc ions. These result raise the possibility that a biosensor can be developed using the lux genes system.

The Role of GADD45 ${\beta}$ in Acute Myocardial Injury (급성 심근 손상에서 GADD45 ${\beta}$의 역할)

  • Cho, Suk-Ki;Hong, Jong-Myeon;Lee, Hak-Mo;Oh, Byong-Chul;Lee, Jae-Woong;Lee, Jeong-Ryul
    • Journal of Chest Surgery
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    • v.41 no.1
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    • pp.25-33
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    • 2008
  • Background: A critical shortage of donor organs has necessitated an investigation of new strategies to increase the availability of additional organs available for human transplantation. We investigated the amount of apoptosis and expression of GADD45 ${\beta}$ in two groups, a GADD45 ${\beta}$-transfected group and untransfected group. Material and Method: The experimental groups consist of a control group (normal H9C2 cell line) and GADD45 ${\beta}$-transfected group. After injury of the each group, we evaluated the expression of GADD45 ${\beta}$ and the level of apoptosis in each group. Result: There was a significant increase in the expression of GADD45 ${\beta}$ in the GADD45 ${\beta}$-transfected group at 1 hour, 2 hours, and 3 hours after stimuli as compared with the control group. The amount of cardiac myoblast cell line apoptosis was significantly lower in the GADD45 ${\beta}$-transfected group as compared with the control group. The concentration of annex in in the GADD45 ${\beta}$-transfected group was significantly lower than that of the control. group after cell. injury. Conclusion: Transfection of a rat myoblast cell line with the GADD45 ${\beta}$ gene results in. decreased susceptibility to cell injury of human serum.

Molecular Cloning of Antagonistic Genes in Pseudomonas maItophiliQ B-14 (토양병해 길항성 Pseudomonas maltophilia B-14의 길항유전자탐색)

  • 구본성;서영우;윤상홍;박경수;은무영;김용환;오상우;류진창;은무영
    • Microbiology and Biotechnology Letters
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    • v.20 no.6
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    • pp.619-624
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    • 1992
  • Tn5 lac 삽입으로 채소입고병원균에 길항력이 약화된 T-67 및 고추역병균과 참깨역병균에 길항력이 약화된 T-81의 Tn5 lac 유전자 일부와 오른쪽 말단에 있는 길항관련 유전자의 flanking sequence가 cloning된 pAG67 및 pAG81 clone을 선발하였고, pAG67 및 pAG81 clone된 길항관련 유전자의 flanking sequence를 야생 길항균 Pseudomonas maltophilia B-14의 DNA를 probe로 사용하여 Southern hybridization으로 확인하였으며, 제한효소 지도를 작성하여 8Kb 및 4Kb 크기의 flanking sequence가 cloning되었음을 확인하였다. pAG6 및 pAG81의 flanking sequence를 EcoRi-BglII와 EcoRI-MpaI으로 분리하여 유전자 은행으로부터 길항관련 유전자가 cloning된 cosmid clone 7개주를 선발하였다.

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An Efficient Algorithm for Determining Probe Specificity in DNA Chips (유전자 칩에서 Probe Specificity를 판별하기 위한 효율적인 알고리즘)

  • Kwon Young-Dae;Park Kyoung-Wook;Lim Hyeong-Seok
    • Proceedings of the Korean Information Science Society Conference
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    • 2005.07a
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    • pp.913-915
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    • 2005
  • 유전자 칩의 정확성은 각 유전자들의 식별자로 활용되는 probe들에 의해 결정된다. 칩에 삽입되는 probe들은 반응오류를 피하기 위해 이중구조, 녹는점, 그리고 CG구조와 같은 요소들을 고려한다. 또한 다른 유전자들과의 교차반응을 최소화하기 위해 specificity를 고려한다. probe의 specificity 검증은 전체 유전자들에 대해 탐색해야 하므로 대규모 염색체에 대해서는 많은 계산이 요구된다. 본 논문에서는 specificity 검증을 위한 효율적인 알고리즘을 제시한다. 제시한 알고리즘은 해시테이블을 활용하여 probe가 specificity를 만족하지 못하게 하는 유전자 시퀀스들만을 탐색하여 비교한다. 제시한 알고리즘이 기존 알고리즘보다 효율적임을 실험결과를 통해 보인다.

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Expression of Glutathione Reductase Gene in Transgenic Tobacco Plant (형질전환 담배 식물체에서 Glutathione Reductase 유전자의 발현)

  • 이효신;조진기
    • Korean Journal of Plant Tissue Culture
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    • v.28 no.2
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    • pp.87-90
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    • 2001
  • BcGRl gene encoding cytosolic glutathione reductase of Chinese cabbage (Brassica campestris var. Pekinensis cv. Seoul) was placed under the control of the CaMV 35S promoter and introduced into tobacco (Nicotiana tabacum L. cv. Samsun) via Agrobacterium-mediated transformation. T$_{0}$ 32 independent plants transformed with BcGRl gene were selected with kanamycin and they were confirmed by polymerase chain reaction (PCR) and Southern blot analysis. Northern blot analysis revealed that the constitutive expression of BcGRl gene and there was no relationship between the copy number of introduced gene and the levels of BcGRl transcripts.

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Molecular Cloning of Escherichia coli cdd Gene Encoding Cytidine/Deoxycytidine Deaminase. (Escherichia coli의 시티딘/디옥시시틴딘 디아미나제를 코드하는 cdd 유전자의 클로닝)

  • 권택규;김태호;황선갑;김종국;송방호;홍순덕
    • Microbiology and Biotechnology Letters
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    • v.18 no.6
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    • pp.640-646
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    • 1990
  • We have cloned the cdd gene from E. coli C600 using (cdd-) as a host. From the sequenced promoter region of E=, coli cdd gene which has been determined by Valentin-Hansen P. (1985), we synthesized the 23 mer oligonucleotides corresponding to the transcription initiation region and used as a probe for cloning of the cdd gene by Southern blotting. The isolated fragments in the blotting were introduced to the colony hybridization after transforming it into the E. coli JF611 (cdd-, pyr double mutant), and we identified the hybridized band at 27 kb long. From the original insert of 27 kb fragment in theBamHI site of pBR322, the 5.3 kb fragment containing the cdd gene was isolated by subsequent deletion and subeloning. From the derived plasmid pTK509, further deletion and subcloning were performed and clarified that the cdd gene was located in the 2.1 kb of SaZI/DraI segment in the insert of pTK605. The polypeptide encoded by the cloned DNA was appeared to be a molecular mass of 33,000.

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Fungal pathogen protection in transgenic lettuce by expression of a apoptosis related Bcl-2 gene (Apoptosis 관련 Bcl-2유전자의 도입을 통한 곰팡이 저항성 형질전환 상추의 육성)

  • Seo, Kyung-Sun;Min, Byung-Whan
    • Journal of Plant Biotechnology
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    • v.38 no.3
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    • pp.209-214
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    • 2011
  • Transgenic lettuce plants were successfully obtained from hypocotyl explants inoculated with Agrobacterium tumefaciens, which harbored a binary vector plasmid with Bcl-2 gene, related to apoptosis. After culture and selection on MS medium a number of kanamycin-resistant plantlets were regenerated. Polymerase chain reaction, Southern blot analysis and Northern blot analysis were used to identify and characterize the transgenic plants with the integrated Bcl-2 gene. Over 100 transgenic plants have been established in soil and flowered in the greenhouse. T1 progeny of 100 transgenic lettuce inbred lines were inoculated with Sclerotinia sclerotiorum. Expression of the Bcl-2 peptide in transgenic lettuce plants provides high levels of field resistance against Sclerotinia sclerotiorum, causal agent of the agronomically important fungal disease of lettuce.

Characteristics of MSX1 gene in Korean nonsyndromic cleft lip and palate individuals (한국인 비증후군성 구순구개열자에서 MSX1 유전자의 특성에 대한 연구)

  • Lee, Hae-Kyung;Kim, Seong-Sik;Son, Woo-Sung
    • The korean journal of orthodontics
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    • v.38 no.2
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    • pp.133-143
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    • 2008
  • Objective: This study was performed to identify the characteristics of the MSX1 gene (locus chromosome 4p16) in Korean nonsyndromic cleft lip and palate (CL/P), which is assumed to be a major candidate gene acting as a causal factor in nonsyndromic CL/P and missing teeth. Methods: The 36 individuals (23 males and 13 females) who had visited the department of orthodontics at from 1998 to 2002 and who had nonsyndromic CL/P were included in the study. Using a PCR-based assay, the MSX1 gene was amplified, sequenced, and searched for inferred protein products (Reference: Homo sapiens MSX1, accession number AF426432 and NP_002439). The common single nucleotide polymorph isms were observed. Results: In exon 1, nucleotide "A" of the 253 basepair (bp) region was substituted for "G", and in the 255 bp region, nucleotide "G" was inserted. In exon 2, nucleotide "C" of the 11 bp region was substituted for "A", and "T" or "G" was inserted into the 351 bp region whereas "T" or "A" was inserted into the 352 bp region. In protein analysis, "Thr85Ala" missense mutation was found. The "Thr85Ala" missense mutation in this study is different from those of studies using subjects of other races. Conclusions: The results suggest that there is specific mutation of MSX1 in Korean and it plays an important role in Korean nonsyndromic CL/P. However, any distinct genetic polymorphisms between CL/P with missing teeth in the cleft region and CL/P without missing teeth could not be found.