• 제목/요약/키워드: DNA mobility

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DNA Heteroduplex Mobility Assay법을 이용한 파이토플라스마 병원체의 유연관계 분석 (Rapid Analysis of Genetic Relationship of Phytoplasma Isolates by a DNA Heteroduplex Mobility Assay)

  • 한상섭;;김성문
    • 한국식물병리학회지
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    • 제14권5호
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    • pp.382-385
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    • 1998
  • Molecular identification and genetic relationships between a phytoplasma associated with chestnut little leaf (CLL) and phytoplasma isolates of other trees in Korea were amplified by polymerase chain reaction (PCR). These 16S rDNA sequences amplified from the various phytoplasmas were used in DNA heteroduplex mobility assays (HMA). In DNA HMA combined with PCR, the mobility shift was observed for a heteroduoplex formed in combined with CLL and jujube witches broom, but not for those formed in combined with CLL and each of sumac witches broom, paulownia witches broom, and mulberry dwarf. HMA combined with PCR has been shown to be a very useful method for detection and differentiation of phytoplasmas.

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Advanced Air Mobility ICT 기술 현황 및 발전 방향 (Current Status and Development Direction of Advanced Air Mobility ICTs)

  • 오봉진;이문수;김법균;정양재;임유진;임채덕
    • 전자통신동향분석
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    • 제38권3호
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    • pp.1-10
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    • 2023
  • In this study, the status of global advanced air mobility (AAM) was investigated to derive information and communications technologies (ICTs) that should be prepared according to directions of domestic AAM development. AAM is an urban air traffic system for moving from city to city by electric vertical take-off and landing or personal aircraft. It is expected to establish a three-dimensional air traffic system that can solve ground traffic congestion caused by the rapid global urbanization. With the full-scale commercialization of AAM solutions, high-density air traffic is expected, and with the advent of the personal air vehicle (PAV), the flight space usage is expected to expand. Therefore, it is necessary to develop a safe AAM service through early research on core ICTs for autonomous flight.

R3V6 Amphiphilic Peptide with High Mobility Group Box 1A Domain as an Efficient Carrier for Gene Delivery

  • Ryu, Jaehwan;Jeon, Pureum;Lee, Minhyung
    • Bulletin of the Korean Chemical Society
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    • 제34권12호
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    • pp.3665-3670
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    • 2013
  • The R3V6 peptide includes a hydrophilic arginine stretch and a hydrophobic valine stretch. In previous studies, the R3V6 peptide was evaluated as a gene carrier and was found to have low cytotoxicity. However, the transfection efficiency of R3V6 was lower than that of poly-L-lysine (PLL) in N2A neuroblastoma cells. In this study, the transfection efficiency of R3V6 was improved in combination with high mobility group box 1A domain (HMGA). HMGA is originated from the nuclear protein and has many positively-charged amino acids. Therefore, HMGA binds to DNA via charge interaction. In addition, HMGA has a nuclear localization signal peptide and may increase the delivery efficiency of DNA into the nucleus. The ternary complex with HMGA, R3V6, and DNA was prepared and evaluated as a gene carrier. First, the HMGA/DNA complex was prepared with a negative surface charge. Then, R3V6 was added to the complex to coat the negative charges of the HMGA/DNA complex, forming the ternary complex of HMGA, R3V6, and DNA. A physical characterization study showed that the ternary complex was more stable than the PLL/DNA complex. The HMGA/R3V6/DNA complex had a higher transfection efficiency than the PLL/DNA, HMGA/DNA, or R3V6/DNA complexes in N2A cells. Furthermore, the HMGA/R3V6/DNA complex was not toxic to cells. Therefore, the HMGA/R3V6/DNA complex may be a useful gene delivery carrier.

Helical Periodicity of $(dT)_n{\cdot}(dA)_n{\cdot}(dT)_n$ Triple - Stranded DNA

  • Kim, Ki-Hyun;Koo, Hyeon-Sook
    • BMB Reports
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    • 제30권6호
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    • pp.426-430
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    • 1997
  • The helical periodicity of the triple-stranded $(dT)_n{\cdot}(dA)_n{\cdot}(dT)_n$ sequence was determined by measuring gel-mobilities of bent DNA fragments containing the sequence. In the bent DNA fragments, a $GA_{22}G$ $CT_{22}C$ sequence was located between two bent DNA loci composed of six $A_{6}{\cdot}T_{6}$ repeats. and the DNA length between the bent DNA loci was varied by 1 base pair over a full helical turn. The gel mobility of each bent DNA fragment reflected the overall extent of DNA bending and varied with the DNA length between the two bent loci. Mobilities of the bent DNA fragments in 5% polyacrylamide gel were measured after preincubating the DNA fragments both in the presence and absence of $CT_{22}C$ oligonucleotide. By comparing the bent DNA fragments containing an intermolecular triplex structure with those of a genuine duplex structure in the gel mobilities, the helical periodicity of the $T_n{\cdot}A_n{\cdot}T_n$ triplex DNA was determined to be $11.5({\pm}0.3)bp/turn$.

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Glycation propagator에 의한 DNA damage 증가 (Increased DNA Damage Induced by Glycation Propagator)

  • 손태건;곽이섭;진영완
    • 생명과학회지
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    • 제14권3호
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    • pp.406-410
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    • 2004
  • Glycation 반응은 glucose와 amino group 간에 일어나는 비효소적 축합 반응인 maillard 반응의 초기 반응으로 non enzymatic glycation 이라고도 한다. 생체내 glycation 반응을 통해 다수의 dicarbonyl화합물이 생성되고, 이들 dicarbonyl들 중에서 매우 반응성이 큰 것으로 확인된 glyoxal과 methylglyoxal과 catalase를 반응 시켜 glycation catalase의 활성 변화를 확인하였다. Non-glycated catalase에 비해 glycation catalase에서 구조적 인 modification과 degradation이 일어났으며, glycation반응 시간에 따라 활성이 크게 저하되는 것으로 확인 할 수 있었다. 특히 glycation 반응 시간 20일 경과 이후 glycation catalase 경우 활성이 거의 상실한 것으로 나타났다 Glyoxal과 methylglyoxal의 농도를 달리 해서 DNA와 반응 시켜 glycation propagator에 의한 직접적인 DNA damage를 확인 한 결과 Glyoxal과 methylglyoxal의 농도와 반응 시간에 따라 DNA mobility sit의 차이를 나타냈다. Fenton reaction 조건에 glyoxal과 methylglyoxal에 의해 활성이 저하된 catalase를 첨가 시켜 8-OH-dG의 생성을 확인한 결과 두 glycation propagator와의 반응 시간 의존적으로 8-OH-dG의 생성이 증가함을 보였다. 이상의 결과를 통해 glyoxal과 methylglyoxal의 antioxidant의 glycation은 oxidative stress의 증사를 유발해 생체내 활성 산소로부터 방어 기작에 심각한 문제를 야기하는 것으로 사료된다.

Binding Aspect of Cyclic AMP Receptor Protein to Symmetrically Synthetic 22-, 28- and 30-Base-Pair lac Promoters

  • Park, Sang-Ho;Lee, Tae-Woo;Hwang, Eun-Suk;Lee, Seung-Ki;Shin, Cha-Gyun;Lee, Bong-Jin
    • 한국자기공명학회논문지
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    • 제1권1호
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    • pp.31-44
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    • 1997
  • The effect of the binding of CRP to the symmetrically synthetic 22, 28, and 30 bp lac promoter was investigated by 1H NMR. The binding of cAMP*CRP to the 22 bp DNA did not bring about any changes in the chemical shift values, but did cause selective line broadening of imino proton resonances of specific base pairs. However, The binding of cAMP*CRP to the 28 and 30 bp DNA brought about large changes on the imino proton resonances that seems to be induced by DNA bending. We studied also the role of cAMP as an activator of DNA/CRP complex formation by gel mobility shift assay. Gel mobility shift assay revealed that the cAMP*CRP complex was not able to bind to the 22 bp DNA fragment, but was able to bind to the 28 bp DNA fragment of lac promoter region.

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대두 ${\beta}-conglycinin$ 유전자 발현의 전사 조절에 관한 연구 -(II) 대두 발달과정 중의 대두 배 인자 3의 역가 변화- (Transcriptional regulation of soybean ${\beta}-conglycinin$ gene expression: -(II) Developmental change of soybean embryo factor 3 activity-)

  • 이경훈;정동효;김우연
    • Applied Biological Chemistry
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    • 제36권6호
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    • pp.553-556
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    • 1993
  • 대두 종자 저장 단백질의 일종인 ${\beta}-conglycinin$${\alpha}'$subunit 유전자 upstream 영역에 결합하여 전사 조절에 관여하리라 추정되는 SEF3(soybean embryo factor 3)의 발현을 조사하기 위하여 대두 핵 추출물을 조제하였다. 두개의 AACCCA를 포함하는 SE3 DNA를 $^{32}P$로 표지한 후 gel mobility shift assay 탐침으로 이용하여 대두 발달 과정 중의 SEF3의 역가를 조사하여 본 결과, 개화 후 16일부터 32일까지의 역가는 증가하나 SE3-SEF3 결합체 이동도는 감소하였다. 핵 추출물을 alkaline phosphatase 처리하면 결합체의 이동도가 다시 증가하였으나 이 현상은 phosphate에 의해 저해되었다. 그리고 결합체의 형성은 반응 pH 6.8과8.5사이에서는 큰 영향을 받지 않았다.

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Expression of Gal4-VP16 and Gal4-DNA binding domain under the control of the T lymphocyte-specific lck proximal promoter in transgenic mice

  • Ryu, Chun-Jeih;Whitehurst, Charles E.;Chen, Jianzhu
    • BMB Reports
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    • 제41권8호
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    • pp.575-580
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    • 2008
  • Thymocyte-specific transcriptional regulatory systems can be used to better understand the relationship between transcription and V(D)J recombination during early T cell development. In this study, we generated transgenic mice expressing the transactivator Gal4-VP16 or the Gal4 DNA binding domain (Gal4-DBD) under the control of the lck proximal promoter, which is only active in immature thymocytes. From these studies Gal4-VP16 and Gal4-DBD expression was shown to significantly alter thymic cellularity and differentiation without significantly changing the $CD3^+$ thymocyte distribution. Furthermore, the presence of Gal4-VP16 or Gal4-DBD in the transgenic thymocytes retarded the mobility of the Gal4 DNA binding motif as determined by a gel mobility shift assay, suggesting that the developmental alteration did not affect the functional property of the transgenic proteins. These results indicated that lck promoter-driven Gal4-VP16 or Gal4-DBD expression did not affect $CD3^+$ mature thymocytes, thus this system can be applied to study transcriptional regulation of transresponder genes in bigenic mouse model thymocytes.

Capillary Gel Electrophoretic Analysis of Cattle Breeds Based on Difference of DNA Mobility of Microsatellite Markers

  • Lee, Mi-Ji;Yoon, Du-Hak;Jeon, Jin-Tae;Eo, Seong-Kug;Kang, Seong-Ho
    • Bulletin of the Korean Chemical Society
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    • 제30권11호
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    • pp.2655-2660
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    • 2009
  • A breed of cattle, i.e., Korean cattle (Hanwoo), was identified based on the DNA mobilities of their microsatellites (MSs) by capillary gel electrophoresis (CGE) with a laser-induced fluorescence (LIF) detector. The MS markers were used for the accurate identification of species-specific genes. The DNA mobilities of the MS markers of Hanwoo and Holstein were measured using a CGE system with a fused-silica capillary (inner diameter of 75 ${\mu}m$, outer diameter of 365 ${\mu}m$, and total length of 50 cm). The capillary was dynamically coated with 1.0% (w/v) polyvinylpyrrolidone ($M_r$ = 1,000,000) and then filled with a mixture of 1.3% (w/v) poly(ethylene oxide) ($M_r$ = 600,000) and 1.9% (w/v) poly(ethylene oxide) (Mr = 8,000,000) as a sieving gel matrix. The species-specific genes of Hanwoo and Holstein were clearly distinguished within 33 min. This CGE assay technique is expected to be a useful analytical method for the fast and accurate identification of breeds of cattle.

Replication origins oriGNAI3 and oriB of the mammalian AMPD2 locus nested in a region of straight DNA flanked by intrinsically bent DNA sites

  • Balani, Valerio Americo;De Lima Neto, Quirino Alves;Takeda, Karen Izumi;Gimenes, Fabricia;Fiorini, Adriana;Debatisse, Michelle;Fernandez, Maria Aparecida
    • BMB Reports
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    • 제43권11호
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    • pp.744-749
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    • 2010
  • The aim of this work was to determine whether intrinsically bent DNA sites are present at, or close to, the mammalian replication origins oriGNAI3 and oriB in the Chinese hamster AMPD2 locus. Using an electrophoretic mobility shift assay and in silico analysis, we located four intrinsically bent DNA sites (b1 to b4) in a fragment that contains the oriGNAI3 and one site (b5) proximal to oriB. The helical parameters show that each bent DNA site is curved in a left-handed superhelical writhe. A 2D projection of 3D fragment trajectories revealed that oriGNAI3 is located in a relatively straight segment flanked by bent sites b1 and b2, which map in previously identified Scaffold/Matrix Attachment Region. Sites b3 and b4 are located approximately 2 kb downstream and force the fragment into a strong closed loop structure. The b5 site is also located in an S/MAR that is found just downstream of oriB.