• 제목/요약/키워드: Data Amplification

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Cloning and Activity Analysis of the FosB Promoter Region from Human Genomic DNA (사람 핵DNA로부터 FosB 유전자 프로모터 클로닝 및 활성도 분석)

  • Na, Han-Heom;Kang, Yoonsung;Kim, Keun-Cheol
    • Journal of Life Science
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    • v.27 no.8
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    • pp.857-863
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    • 2017
  • The FBJ murine osteosarcoma viral oncogene homolog B (FosB) gene is located at chromosome 19, and encodes 43 Kda protein. Functionally, the FosB gene is important for differentiation, development, and pathogenesis. Furthermore, the FosB gene is suggested as possible biomarker for tracing disease prognosis. In this study, we constructed plasmid containing a FosB promoter region and evaluate its promoter activity. We analyzed the putative promoter region in FosB genomic DNA using bioinformatics program, and we found important regulatory elements in 1 Kb upstream from transcription start site (TSS). Therefore, we performed polymerase chain reaction (PCR) amplification on region from-1,555 upstream to +73 of the FosB genomic DNA, and PCR product was inserted into TA vector to create the $TA-1^{st}FosBp$ plasmid. We then prepared the primer sets, which contain a restriction enzyme site for Kpn1 and Nhe1, in order to reinsert into the TA vector to prepare $TA-2^{nd}FosBp$ plasmid. It was finally subcloned into pGL3-luc vector after enzyme cutting. To evaluate whether the cloned plasmid is useful in cell based experiment, we performed luciferase assay with pGL3-FosBp-luctransfection. FosB promoter activity was increased compared to empty vector, and this activity was significantly increased by treatment of doxorubicin and taxol. We obtained consistent data on regulation of FosB gene expression after anticancer drug treatment using Western blot analysis. The results suggest that promoter cloning of the human FosB gene is very useful for studying gene expression and analyzing biomarkers.

Prevalence of Trichomonas vaginalis by PCR in Men Attending a Primary Care Urology Clinic in South Korea

  • Seo, Jun-Hyeok;Yang, Hye-Won;Joo, So-Young;Song, Su-Min;Lee, Yu-Ran;Ryu, Jae-Sook;Yoo, Eun Sang;Lee, Won Kee;Kong, Hyun-Hee;Lee, Sang-Eun;Lee, Won-Ja;Goo, Youn-Kyoung;Chung, Dong-Il;Hong, Yeonchul
    • Parasites, Hosts and Diseases
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    • v.52 no.5
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    • pp.551-555
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    • 2014
  • Trichomonas vaginalis, a causative agent of trichomoniasis, may trigger symptomatic or asymptomatic non-gonococcal urethritis and chronic prostatitis in men. Despite the availability of highly sensitive diagnostic tests, such as nucleic acid amplification tests, including PCR, few prospective studies present data on male T. vaginalis infection in South Korea. In the present study, the prevalence of T. vaginalis and associated clinical conditions were evaluated in 201 male patients from a primary care urology clinic in South Korea. The prevalence of T. vaginalis infection in our cohort was 4% (8/201) by PCR. T. vaginalis infection was common in men older than 40 years (median age, 52 years). Among the 8 Trichomonas-positive patients, 87.5% (7/8) had prostatic diseases, such as prostatitis and benign prostatic hyperplasia, and 25.0% (2/8) and 12.5% (1/8) were coinfected with Chlamydia trachomatis and Mycoplasma genitalium, respectively. Our results suggest that T. vaginalis infection is not rare in men attending primary care urology clinics in South Korea, especially in those older than 40 years, in whom it may explain the presence of prostatic disease. The possibility of T. vaginalis infection should be routinely considered in older male patients with prostatic diseases in South Korea.

Analysis of Genetic Relatedness by Random Amplified Polymorphic DNA (RAPD) in Pecan Taxa (RAPD를 이용한 Pecan 품종의 유전적 관계 분석)

  • 신동영;김회택;박종인;노일섭
    • Korean Journal of Plant Resources
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    • v.13 no.1
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    • pp.1-10
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    • 2000
  • Pecan is deciduous tree and belongs to the Julandaceae family. Pecan is an economically important as a nut and timber crop. Heterozygosity is expected to be high for typically cross-pollinated. Yet little is known about the nature of genetic variation within this species. In addition, the pedigree of many pecan cultivars remains unknown or is questionable. In this study, the phylogenetic relationships between 22 pecan cultivars and its analyzed by RAPD (randomly amplified polymorphic DNA). PCR Amplification used 40 randomly selected oligoes as primers. Based on their genetic similarities derived from the RAPD data, the 22 pecan cultivars were classified into different five groups in agarose gel. The 22 pecan cultivars were classified into five sectional groups by UPGMA clustering analysis, too. C. flacra and Black walnut showed the 0.9 of similarity index and Farley, Pawnee showed the 0.85 of similarity index. The 22 pecan cultivars were classified into different five groups by analysis of the 4% polyacrylamide gel fraction. (Group I : 1, 2, 3, 4, 13, 16, 17, 20, 21 Group II : 14,18 GroupIII : 6,12 GroupIV : 5, 11, 15, 19, 22 CroupV : 7, 8, 9, 10) Group V show the 1.0 of similarity index and Farley, Sturya, Clarke, Pawnee show the 0.98 of similarity index and Kiowa, Schley show the 0.92 of similarity index. Results from this study indicated that RAPD can be used to establish the genetic relationships among the 22 pecan cultivars. Similarity coefficients generally agreed with what would be predicted in cultivars with known pedigrees, and we could accurately construct relationships among cultivars. In addition, we have shown that RAPD provides useful information on the origin of unknown cultivars.

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Identification of Phellinus linteus by Morphological Characteristics and Molecular Analysis (형태적.분자생물학적 방법에 의한 Phellinus linteus의 동정에 관한 연구)

  • Kim, Sang-Hee;Kim, Soo-Ho;Sung, Jae-Mo;Harrington, Thomas C.
    • The Korean Journal of Mycology
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    • v.27 no.5 s.92
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    • pp.337-340
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    • 1999
  • The context and upper surface of Phellinus basidiocarp become blackened, rimose and woody. The basidiocarp is sessile, dimidiate and elongate. The basidiospores are pigmented and ovoid to globose. Hymenial setae are $17{\sim}35{\times}6{\sim}8{\mu}m$. Nineteen isolates of Phellinus species, including Phellinus linteus, were used for sequencing of the internal transcribed spacer (ITS) region of the nuclear rDNA. Based on these sequence data, specific primers were designed for identification of Phellinus linteus isolates in Korea. The specific primers were within the ITS1 and ITS2 regions and were nested within the universal primers flanking the spacer regions. A total of four primers (the universal primers ITS-1F and ITS-4, and the specific primers PL-F and PL-R) were used for detection of Phellinus linteus collected in Korea. The length of the four amplification products of Phellinus linteus DNA were 800 bp (ITS-1F/ITS-4), two bands of about 720 bp (ITS-1F/PL-R and PL-F/ITS-4), and 610 bp (PL-F/PL-R). Among 23 isolates of Phellinus species collected in Korea, Thirteen isolates were identified as Phellinus linteus based on the presence of the four bands. The other species produced only the single ITS-1F/ITS-4 product.

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Mutation Analysis of Synthetic DNA Barcodes in a Fission Yeast Gene Deletion Library by Sanger Sequencing

  • Lee, Minho;Choi, Shin-Jung;Han, Sangjo;Nam, Miyoung;Kim, Dongsup;Kim, Dong-Uk;Hoe, Kwang-Lae
    • Genomics & Informatics
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    • v.16 no.2
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    • pp.22-29
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    • 2018
  • Incorporation of unique barcodes into fission yeast gene deletion collections has enabled the identification of gene functions by growth fitness analysis. For fine tuning, it is important to examine barcode sequences, because mutations arise during strain construction. Out of 8,708 barcodes (4,354 strains) covering 88.5% of all 4,919 open reading frames, 7,734 barcodes (88.8%) were validated as high-fidelity to be inserted at the correct positions by Sanger sequencing. Sequence examination of the 7,734 high-fidelity barcodes revealed that 1,039 barcodes (13.4%) deviated from the original design. In total, 1,284 mutations (mutation rate of 16.6%) exist within the 1,039 mutated barcodes, which is comparable to budding yeast (18%). When the type of mutation was considered, substitutions accounted for 845 mutations (10.9%), deletions accounted for 319 mutations (4.1%), and insertions accounted for 121 mutations (1.6%). Peculiarly, the frequency of substitutions (67.6%) was unexpectedly higher than in budding yeast (~28%) and well above the predicted error of Sanger sequencing (~2%), which might have arisen during the solid-phase oligonucleotide synthesis and PCR amplification of the barcodes during strain construction. When the mutation rate was analyzed by position within 20-mer barcodes using the 1,284 mutations from the 7,734 sequenced barcodes, there was no significant difference between up-tags and down-tags at a given position. The mutation frequency at a given position was similar at most positions, ranging from 0.4% (32/7,734) to 1.1% (82/7,734), except at position 1, which was highest (3.1%), as in budding yeast. Together, well-defined barcode sequences, combined with the next-generation sequencing platform, promise to make the fission yeast gene deletion library a powerful tool for understanding gene function.

PREVALENCE OF STREPTOCOCCUS MUTANS AND STREPTOCOCCUS SOBRINUS IN CHILDREN WITH MIXED DENTITION (혼합치열기 어린이에서 Streptococcus mutans와 Streptococcus sobrinus의 분포도 조사)

  • Lee, Myung-Sung;Choi, Sung-Chul;Park, Jae-Hong
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.2
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    • pp.247-254
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    • 2007
  • Mutans streptococci have been reported to be implicated in dental caries. Of these streptococcal species, Streptcoccus mutans and Streptococcus sobrinus are most commonly found in human dental caries. Prevalence of these bacterial species in dental caries is found to be varied in different races and countries. Therefore, importance of these bacteria in dental caries remains to be determined The present study was performed to find out correlation S. mutans and S. sobrinus with dental caries in 125 Korean children with mixed dentition between 6 to 11 years of age. They were classified as group A(6-8 years) and group B(9-11 years) by age. For the study, stimulated saliva samples were collected from each subject. The vials containing saliva specimens were serially diluted (1:10) in saline and plated in duplicate on tryptone-yeast extract-cysteine with sucrose and bacitracin (TYCSB) for S. mutans and S. sobrinus. After genomic DNA was extracted from the samples, polymerase chain reaction (PCR) amplification was performed for identification using universal primers and specific primers to S. mutans and S. sobrinus. Data of microbial variables were compared to caries status of the subjects. According to this study, the result were as follows : 1. S. mutans versus S. sobrinus were moderate positive linear correlated in both group A(r=0.70) and group B(r=0.50). 2. Between S. mutans and dental caries there were weak positive linear correlation in both group A(r= 0.25) and group B(r=0.34). 3. S. sobrinus versus dental caries were not correlated in group A but slightly correlated in group B(r=0.21). 4. Between S. mutans and age, there were not correlation in both group. 5. S. sobrinus versus age were weak correlated in group A(r=0.32) but not correlated in group B.

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Anti-proliferative Effects of Atractylis lancea (Thunb.) DC. via Down-regulation of the c-myc/hTERT/Telomerase Pathway in Hep-G2 Cells

  • Guo, Wei-Qiang;Li, Liang-Zhi;He, Zhuo-Yang;Zhang, Qi;Liu, Jia;Hu, Cui-Ying;Qin, Fen-Ju;Wang, Tao-Yun
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.11
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    • pp.6363-6367
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    • 2013
  • Atractylis lancea (Thunb.) DC. (AL), an important medicinal herb in Asia, has been shown to have anti-tumor effects on cancer cells, but the involved mechanisms are poorly understood. This study focused on potential effects and molecular mechanisms of AL on the proliferation of the Hep-G2 liver cancer cell line in vitro. Cell viability was assessed by MTT test in Hep-G2 cells incubated with an ethanol extract of AL. Then, the effects of AL on apoptosis and cell cycle progression were determined by flow cytometry. Telomeric repeat amplification protocol (TRAP) assays was performed to investigate telomerase activity. The mRNA and protein expression of human telomerase reverse transcriptase (hTERT) and c-myc were determined by real-time RT-PCR and Western blotting. Our results show that AL effectively inhibits proliferation in Hep-G2 cells in a concentrationand time-dependent manner. When Hep-G2 cells were treated with AL after 48h,the $IC_{50}$ was about 72.1 ${\mu}g/mL$. Apoptosis was induced by AL via arresting the cells in the G1 phase. Furthermore, AL effectively reduced telomerase activity through inhibition of mRNA and protein expression of hTERT and c-myc. Hence, these data demonstrate that AL exerts anti-proliferative effects in Hep-G2 cells via down-regulation of the c-myc/hTERT/telomerase pathway.

Smartphone Usage Experience of College students (대학생들의 스마트폰 사용 경험)

  • Kim, Jungae;Cho, Eui-young
    • The Journal of the Institute of Internet, Broadcasting and Communication
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    • v.16 no.3
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    • pp.187-201
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    • 2016
  • The purpose of this study was to analyze the smart phone usage experience structure for college students. The phenomenology approach was used by conducting 4 times in-depth interviews with the 7 college students age between 20~23. Collected Data was analyzed by Giorgi's phenomenological method. There were 4 general structure descriptions and 28 themes. General structure descriptions were 'Dependence on smart phone', 'Phenomenon of smart phone usage', 'Results of smart phone usage' and 'Relationship between stress and smart phone'. Themes consisted of 'Ambivalence', 'Positive feelings for smart phones', 'Separation anxiety', 'Negative feelings for smart phones'. 'Dependent feelings for smart phones', 'A sense of unity with smart phones', 'Helpful for my life', 'Deeply involved with my life', 'Useful tool in my life', 'Study, communication, relaxation and health care', 'Maintain personal relationship', 'The most important thing in my life', 'Shallow relationship stress', 'Interference with personal relationship', 'Forming a negative relationship', 'Invasion of privacy', 'Forming a Incorrect relationship', 'Unwanted participation', 'No effort to improve relationship', 'Poor concentration', 'Interference with academic study', 'Study assistance', 'Limited help for study in depth', 'Shelter from the stress', 'Amplification of the stress', 'Obsessive focus', 'Feel separation for the reality' and 'Waste of time'. Smart phone usage make college students dependent on smart phones and had negative influence on both personal relationship and academic results. Usage of smart phone also amplified stress. Therefore we need fundamental principles on usage of smart phone and propose to build cultural etiquette that provides reasonable way to use smartphone.

Development of depression diagnosis system using EEG signal (뇌파 측정 신호를 이용한 우울증 진단장치 개발)

  • Kim, Kyu-Sung;Jung, Ju-Hyeon;Lee, Woo-Cheol
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.12
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    • pp.452-458
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    • 2017
  • In this study, a device was developed for diagnosing depression using EEG signals from July 2016 to June 2017. For normal people, the left alpha rhythm is more activated than the right alpha rhythm, but for the depressed patients, the right alpha rhythm is more activated than the left one. An analog circuit and digital low pass filter were used for noise removal and amplification of EEG, and the Hamming window function was applied to eliminate the signal leakage generated by the fast Fourier transform. To verify the validity of the developed diagnosis system, the EEG of 20 university students in the 3rd and 4th grade with an average age of 24 years was measured. Calculations of the relative value of the left and right alpha rhythm for the depression diagnosis revealed a minimum, maximum, and mean value of 66.7, 113.3, and 92.2, respectively. In addition, 7 out of 20 subjects were between 90 and 95, and those with a higher mean deviation of approximately 20 tended to have mild depression. These results can provide meaningful data for the development of depression treatment equipment by solving the left and right brain asymmetry problem, and it may be applied usefully to diagnose depression after clinical trials on a large number of depressed patients.

Prevalence and Genetic Characterization of Toxoplasma gondii in House Sparrows (Passer domesticus) in Lanzhou, China

  • Cong, Wei;Huang, Si-Yang;Zhou, Dong-Hui;Zhang, Xiao-Xuan;Zhang, Nian-Zhang;Zhao, Quan;Zhu, Xing-Quan
    • Parasites, Hosts and Diseases
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    • v.51 no.3
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    • pp.363-367
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    • 2013
  • The prevalence of Toxoplasma gondii infection in birds has epidemiological significance because birds are indeed considered as a good indicator of environmental contamination by T. gondii oocysts. In this study, the prevalence of T. gondii in 313 house sparrows in Lanzhou, northwestern China was assayed by the modified agglutination test (MAT). Antibodies to T. gondii were positive in 39 (12.46%) of 313 samples (MAT titer ${\geq}$ 1:5). Tissues of heart, brain, and lung from the 39 seropositive house sparrows were tested for T. gondii DNA, 11 of which were found to be positive for the T. gondii B1 gene by PCR amplification. These positive DNA samples were typed at 9 genetic markers, including 8 nuclear loci, i.e., SAG1, 5'- and 3'-SAG2, alternative SAG2, SAG3, GRA6, L358, PK1, c22-8 and an apicoplast locus Apico. Of them, 4 isolates were genotyped with complete data for all loci, and 2 genotypes (Type II variants; ToxoDB #3 and a new genotype) were identified. These results showed that there is a potential risk for human infection with T. gondii in this region. To our knowledge, this is the first report of T. gondii seroprevalence in house sparrows in China.