• Title/Summary/Keyword: ms DNA

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Value of PAX1 Methylation Analysis by MS-HRM in the Triage of Atypical Squamous Cells of Undetermined Significance

  • Li, Shi-Rong;Wang, Zhen-Ming;Wang, Yu-Hui;Wang, Xi-Bo;Zhao, Jian-Qiang;Xue, Hai-Bin;Jiang, Fu-Guo
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.14
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    • pp.5843-5846
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    • 2015
  • Background: Detection of cervical high grade lesions in patients with atypical squamous cells of undetermined significance (ASCUS) is still a challenge. Our study tested the efficacy of the paired boxed gene 1 (PAX1) methylation analysis by methylation-sensitive high-resolution melting (MS-HRM) in the detection of high grade lesions in ASCUS and compared performance with the hybrid capture 2 (HC2) human papillomavirus (HPV) test. Materials and Methods: A total of 463 consecutive ASCUS women from primary screening were selected. Their cervical scrapings were collected and assessed by PAX1 methylation analysis (MS-HRM) and high-risk HPV-DNA test (HC2). All patients with ASCUS were admitted to colposcopy and cervical biopsies. The Chisquare test was used to test the differences of PAX1 methylation or HPV infection between groups. Results: The specificity, sensitivity, and accuracy for detecting CIN2 + lesions were: 95.6%, 82.4%, and 94.6%, respectively, for the PAX1 MS-HRM test; and 59.7%, 64.7%, and 60.0% for the HC2 HPV test. Conclusions: The PAX1 methylation analysis by MS-HRM demonstrated a better performance than the high-risk HPV-DNA test for the detection of high grade lesions (CIN2 +) in ASCUS cases. This approach could screen out the majority of low grade cases of ASCUS, and thus reduce the referral rate to colposcopy.

Genetic Transformation of Sweet Potato by Particle Bombardment (Particle Bombardment에 의한 고구마의 형질전환)

  • 민성란;정원중;이영복;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.329-333
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    • 1998
  • $\beta$-Glucuronidase (GUS) gene of Escherichia coli was introduced into sweet potato (Ipomoea batatas (L.) Lam.) cells by particle bombardment and expressed in the regenerated plants. Microprojectiles coated with DNA of a binary vector pBI121 carrying CaMV35S promoter-GUS gene fusion and a neomycin phosphotransferase gene as selection marker were bombarded on embryogenic calli which originated from shoot apical meristem-derived callus and transferred to Murashige and Skoog (MS) medium supplemented with 1 mg/L 2,4-dichlorophenoxyacetic acid and 100 mg/L kanamycin. Bombarded calli were subcultured at 4 week intervals for six months. Kanamycin-resistant calli transferred to MS medium supplemented with 0.03 mg/L 2iP, 0.03 mg/L ABA, and 50 mg/L kanamycin gave rise to somatic embryos. Upon transfer to MS basal medium without kanamycin, they developed into plantlets. PCR and northern analyses of six regenerants transplanted to potting soil confirmed that the GUS gene was inserted into the genome of the six regenerated plants. A histochemical assay revealed that the GUS gene was preferentially expressed in the vascular bundle and the epidermal layer of leaf, petiole, and tuberous root.

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Electroporation Conditions for DNA Transfer into Somatic Embryogenic Cells of Zoysia japonica (들잔디 체세포 배발생 세포로의 DNA 전입을 위한 Electroporation 조건 구명)

  • 박건환;안병준
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.1
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    • pp.13-19
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    • 1998
  • We have reported previously that intact embryogenic cells can be used instead of protoplasts for electroporation-mediated transformation of zoysiagrass and rice. In this study, conditions of the tissue electroporation were examined to optimize the procedures. Embryogenic cell suspensions were established in liquid MS medium containing 2 mg/L of 2,4-D with embryogenic calluses induced from mature embryos of Z. japonica. The suspension-cultured cell clumps were electroporated with 35S-gusA expression vector DNA, and degrees of DNA introduction into the cells were determined by histological expression rates of the gusA marker gene. DNA transfer into the cell clumps occurred in wide range of voltage (100-400 V) and capacitance (10-1980 $\mu\textrm{F}$), but more in the ranges of 200-300 V and 330-800 $\mu\textrm{F}$ DNA concentrations higher than 6 $\mu\textrm{g}$/mL were adequate for GUS expression of the electroporated cells. DNA transfers were confirmed in all three embryogenic cell lines but only in one out of eleven non-embryogenic lines. Positive GUS expressions occurred with DNAs added even 20-40 h after pulse treatments. As a promoter of gusA, Act1 and Ubi1 were effective 7 and 5 times than 35S respectively in number of GUS expression units on electroporated cell clumps. Embryogenic cell clumps survived and regenerated into plantlets after pulse treatments of wide range of conditions.

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Restriction Fragment Length Polymorphisms of Genomic DNA in Strains of Xanthomonas campestris pv. vesicatoria (지리적 기원이 다른 고추 더뎅이병균 균주 Genomic DNA의 RFLP 분석)

  • 정희정
    • Korean Journal Plant Pathology
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    • v.12 no.2
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    • pp.162-168
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    • 1996
  • 우리 나라의 주요 고추 재배지와 미국, 대만, 호주, 아르헨티나에서 수집된 44 개 고추 더뎅이병균(Xanthomonas campestris pv. vesicatoria)균주간의 유전적변이를 genomic DNA의 restriction fragment length polymorphism(RFLP)에 의해 분석하였다. Genomic DNA RFLP profiles을 cluster 분석하여 얻은 dendrogram에서 지리적 기원이 다른 44개 균주들은 11개 RFLP 그룹으로 분류되었다. 외국 균주들은 genomic DNA의 RFLP 분석에 의해 모두 각각 다른 RFLP 그룹으로 분류되었다. 외국 균주들 중에서 미국 균주는 우리 나라 일부 균주들과 밀접한 유전적 관련성을 가지고 함께 cluster를 이루었는데, 이것은 이 균주들이 동일한 고추 더뎅이병균의 조상 균주 집단에서 유래했으리라는 것을 시사해 준다. 우리 나라 균주들은 6개의 RFLP 그룹으로 분류되었다. 대부분의 우리 나라 균주들은 가까운 cluster를 이루며 미국 균주를 제외한 외국 균주들과 뚜렷하게 구분되었다. 그러나 우리 나라 균주들 중에서 마산에서 수집된 Ms93-1은 다른 우리 나라 균주들과 뚜렷하게 구분되었다. 유전적으로 격리된 균주의 출현은 우리 나라에서 지리적 기원이 다른 고추 더뎅이병균 균주 사이에 이미 발생한 다양한 유전적 분화의 결과라고 추론된다.

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Characterization of Plants Induced by in vitro Culture of Leaf Blade-segments in a Variegated Tobacco (Nicotiana tabacum L. cv. BY-4) (Variegated 담배 (Nicotiana tabacum L. cv. BY-4)의 잎 절편 배양에 따른 재생 식물체의 특성)

  • 배창휴;이효연
    • Korean Journal of Plant Tissue Culture
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    • v.26 no.4
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    • pp.245-250
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    • 1999
  • Plantlets derived from leaf blade-segment culture of a variegated tobacco (Nicotiana tabacum L. cv. BY-4) that was induced by a heavy-ion ($^{14}N$) beam irradiation to proembryos, were characterized. When explants from both white and green sections of leaves of the variegated plant were cultured on MS medium containing 0.1 mg/L NAA and 1.0 mg/L BAP, the white sections yielded only white shoots, whereas the green sections generated approximately 47.2% green, 37.4% white and 15.4% variegated shoots. In the F1 generation of a green tobacco derived from the leaf blade-segment culture, the segregation ratio of green to white was 1,651:54. Furthermore, reciprocal crosses showed that all of the progenies was green, indicating that the variegation is not maternally inherited. When the signal intensity of photosynthesis genes was determined by DNA gel blot analysis using the variegated leaves derived from green sections of variegated leaves, there were more of the rbcL, psbA, 16S rDNA and 23S rDNA chloroplast genes in the white sections than the chloroplast genes in wild type and green sections of the variegated plants.

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Depurination of Nucleosides and Calf Thymus DNA Induced by 2-Bromopropane at the Physiological Condition

  • Sherchan, Jyoti;Choi, Ho-Young;Lee, Eung-Seok
    • Bulletin of the Korean Chemical Society
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    • v.30 no.10
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    • pp.2309-2317
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    • 2009
  • Depurination, the release of purine bases from nucleic acids by hydrolysis of the N-glycosidic bond, gives rise to alterations of the cell genome. Though cells have evolved mechanisms to repair these lesions, unrepaired apurinic sites have been shown to have two biological consequences: lethality and base substitution errors. 2-Bromopropane (2-BP) is used as an intermediate in the synthesis of pharmaceuticals, dyes, and other organics. In addition, 2-BP has been used as a replacement for chloroflurocarbons and 1,1,1-trichloroethane as a cleaning solvent in electronics industry. However, 2-BP was found to cause reproductive and hematopoietic disorders in local workers exposed to it. Owing to the toxicity of 2-BP, there has been a tendency to use 1-BP as an alternative cleaning solvent to 2-BP. However, 1-BP has also been reported to be neurotoxic in rats. Though $N^7$-guanine adduct of 2-BP has been reported previously, massive depurination of the nucleosides and calf thymus DNA was observed in this study. We incubated the nucleosides (ddG, dG, guanosine, ddA, dA and adenosine) with excess amount 2-BP at the physiological condition (pH 7.4, $37\;{^{\circ}C}$), which were analyzed by HPLC and LC-MS/MS. In addition, the time and dose response relationship of depurination in nucleosides induced by 2-bromopropane at the physiological condition was investigated. Similarly, incubation of calf-thymus DNA with the excess amount 2-BP at the physiological condition was also performed. In addition, the time and dose response relationship of depurination in calf-thymus DNA induced by 2-BP at the physiological condition was investigated. Those results suggest that the toxic effect of 2-BP could be both from the depurination of nucleosides and DNA adduct formation.

Genetic Distances of Rainbow Trout and Masu Salmon as Determined by PCR-Based Analysis

  • Yoon, Jong-Man
    • Development and Reproduction
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    • v.24 no.3
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    • pp.241-248
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    • 2020
  • This study used a PCR-based genetic analysis platform to create a hierarchical polar dendrogram of Euclidean genetic distances for two salmonid species, Oncorhynchus mykiss (rainbow trout, RT) and Oncorhynchus masou (masu salmon, MS). The species were distantly related to other fish species based on PCR results from using the designed oligonucleotide primer series. Five oligonucleotide primers were used to generate 330 and 234 scorable fragments in the RT and MS populations, respectively. The DNA fragments ranged in size from approximately 50 bp to more than 2,000 bp. The bandsharing (BS) results showed that the RT population had a higher average BS value (0.852) than that for the MS population (0.704). The genetic distance between individuals supported the presence of adjacent affiliation in cluster I (RT 01-RT 11). The observation of a significant genetic distance between the two Oncorhynchus species verifies that this PCR-based technique can be a useful approach for individual- and population-based biological DNA investigations. The results of this type of investigation can be useful for species safekeeping and the maintenance of salmonid populations in the mountain streams of Korea.

Analysis of Minisatellite 7 of SLC6A19 (SLC6A19-MS7) for the Relationship to Myocardial Infarction and Evolutional Level (SLC6A19 Minisatellites 7(SLC6A19-MS7)의 심근경색과의 관련성과 진화적 분석)

  • Seol, So-Young;Lee, Sang-Yeop;Yum, Ji-Hoon;Yoon, Hae-Soon;SunWoo, Yang-Il
    • YAKHAK HOEJI
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    • v.54 no.1
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    • pp.49-54
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    • 2010
  • SLC6A19 which reported as a neurotransmitter was composed of seven minisatellites. In previous our study, the minisatellites variants of SLC6A19-MS7 showed the susceptibility for hypertension. When this minisatellte sequences were analyzed using the bioinformatic tool, USF1 (upstream transcription factor 1) was found in this region as a putative transcription factor binding site. USF1 is binding with E-boxes which has a consensus sequence of CACGTG. USF1 is a ubiquitously expressed transcription factor and involved in the transcriptional control of many genes including the molecular pathogenesis of cardiovascular disease. Thus, we investigated that the putative functional relationship between the minisatellites variants and susceptibility for myocardial infarction. A case-control study was performed that compared genomic DNA from 400 controls and 225 cases with myocardial infarction. There were no significant differences observed in the overall allelic distribution of minisatellites between controls and cases, which indicates that this polymorphism is not responsible for myocardial infarction susceptibility. Hence, we analyzed the five different minisatellites alleles from this study and characterized 14 different repeats units (Unit1~Unit14). Then, we evaluated the DNA composition, phylogenic tree, and pairwise distances of its repeats. The variability of each repeats differed from 2.33% to 16%. The phylogenic trees for the four SLC6A19-MS7 minisatellites exhibited very different shapes in their braches and distances, and present most common 8 repeats allele was the longest 14 repeats allele. Therefore, this result may help to understand for the evolutional level of the length of minisatellites.

Expression of Chinese Cabbage Glutathione Reductase Gene in Lettuce (Lactuca sativa L.) (형질전환 상추에서 배추 Glutathione Reductase 유전자의 발현)

  • 정재동;김창길;조진기
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.4
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    • pp.267-271
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    • 1998
  • Cotyledon explants of lettuce were cocultured with Agrobacterium tumefaciens LBA4404::pBKS-GR1 harboring glutathione reductase(GR) gene in MS medium supplemented with 0.1 mg/L NAA and 1.0 mg/L 2ip for 48 hr. These explants were transferred to MS medium supplemented with 0.1 mg/L NAA and 1.0 mg/L 2ip, 50 mg/L kanamycin, and 500 mg/L carbenicillin. After 4 weeks of subculture, kanamycin-resistant shoots were obtained on selection medium. Leaves of putative transformants survived on selection medium containing 100 mg/L kanamycin. Incoporation of the GR gene into lettuce was confirmed by PCR analysis of genomic DNA. Southern blot analysis showed that ECL-labeled GR gene was hybridized to the expected amplified genomic DNA fragment of about 1.8 kb from transgenic lettuce. The presence of mRNA in transgenic lettuce was confirmed by RT-PCR with total RNA of transgenic lettuce. In progeny test of transformants, R$_1$ seeds were resistant to kanamycin (200mg/L) on MS medium.

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Looking through the Mass-to-Charge Ratio: Past, Present and Future Perspectives

  • Shin, Seung Koo
    • Mass Spectrometry Letters
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    • v.12 no.4
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    • pp.126-130
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    • 2021
  • The mass spectrometry (MS) provides the mass-to-charge ratios of atoms, molecules, stable/metastable complexes, and their fragments. I have taken a long journey with MS to address outstanding issues and problems by experiments and theory and gain insights into underlying principles in chemistry. By looking through the mass-to-charge ratio, I have studied thermochemical problems in silicon chemistry, the infrared multiphoton dissociation spectroscopy of organometallic intermediates, unimolecular dissociations of halotoluene radical cations, and the kinetics of association/dissociation of alkali halide triple ions with Lewis bases. Various MS platforms have been used to characterize non-covalent interactions between porphyrins and fullerenes and those between the group IIB ions and trioctylchalcogenides, and to examine the binding of the group IA, IIA and porphyrin ions to G-quadruplex DNA. Recently, I have focused on mass-balanced H/D isotope dipeptide tags for MS-based quantitative proteomics, a simple chemical modification method for MS-based lipase assay, and the kinetics and dynamics of energy-variable collision-induced dissociation of chemically modified peptides. Now, I see an important role of MS in global issues in the post-COVID era, as the society demands high standards for indoor air quality to contain the airborne-pathogen transmission as well as in-situ monitoring and tracking of carbon emissions to reduce global warming.