• Title/Summary/Keyword: Genome Analysis

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Highly efficient production of transgenic Scoparia dulcis L. mediated by Agrobacterium tumefaciens: plant regeneration via shoot organogenesis

  • Aileni, Mahender;Abbagani, Sadanandam;Zhang, Peng
    • Plant Biotechnology Reports
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    • v.5 no.2
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    • pp.147-156
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    • 2011
  • Efficient Agrobacterium-mediated genetic transformation of Scoparia dulcis L. was developed using Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pCAMBIA1301 with ${\beta}$-glucuronidase (GUS) (uidA) and hygromycin phosphotransferase (hpt) genes. Two-day precultured leaf segments of in vitro shoot culture were found to be suitable for cocultivation with the Agrobacterium strain, and acetosyringone was able to promote the transformation process. After selection on shoot organogenesis medium with appropriate concentrations of hygromycin and carbenicillin, adventitious shoots were developed on elongation medium by twice subculturing under the same selection scheme. The elongated hygromycin-resistant shoots were subsequently rooted on the MS medium supplemented with $1mg\;l^{-1}$ indole-3-butyric acid and $15mg\;l^{-1}$ hygromycin. Successful transformation was confirmed by PCR analysis using uidA- and hpt-specific primers and monitored by histochemical assay for ${\beta}$-GUS activity during shoot organogenesis. Integration of hpt gene into the genome of transgenic plants was also verified by Southern blot analysis. High transformation efficiency at a rate of 54.6% with an average of $3.9{\pm}0.39$ transgenic plantlets per explant was achieved in the present transformation system. It took only 2-3 months from seed germination to positive transformants transplanted to soil. Therefore, an efficient and fast genetic transformation system was developed for S. dulcis using an Agrobacterium-mediated approach and plant regeneration via shoot organogenesis, which provides a useful platform for future genetic engineering studies in this medicinally important plant.

Functional analysis of SH3 domain containing ring finger 2 during the myogenic differentiation of quail myoblast cells

  • Kim, Si Won;Lee, Jeong Hyo;Park, Tae Sub
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.8
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    • pp.1183-1189
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    • 2017
  • Objective: Owing to the public availability of complete genome sequences, including avian species, massive bioinformatics analyses may be conducted for computational gene prediction and the identification of gene regulatory networks through various informatics tools. However, to evaluate the biofunctional activity of a predicted target gene, in vivo and in vitro functional genomic analyses should be a prerequisite. Methods: Due to a lack of quail genomic sequence information, we first identified the partial genomic structure and sequences of the quail SH3 domain containing ring finger 2 (SH3RF2) gene. Subsequently, SH3RF2 was knocked out using clustered regularly interspaced short palindromic repeat/Cas9 technology and single cell-derived SH3RF2 mutant sublines were established to study the biofunctional activity of SH3RF2 in quail myoblast (QM7) cells during muscle differentiation. Results: Through a T7 endonuclease I assay and genotyping analysis, we established an SH3RF2 knockout (KO) QM7#4 subline with 61 and 155 nucleotide deletion mutations in SH3RF2. After the induction of myotube differentiation, the expression profiles were analyzed and compared between regular QM7 and SH3RF2 KO QM7#4 cells by global RNA sequencing and bioinformatics analysis. Conclusion: We did not detect any statistically significant role of SH3RF2 during myotube differentiation in QM7 myoblast cells. However, additional experiments are necessary to examine the biofunctional activity of SH3RF2 in cell proliferation and muscle growth.

Uridylate kinase as a New Phylogenetic Molecule for Procaryotes

  • Lee, Dong-Geun;Lee, Jin-Ok;Lee, Jae-Hwa
    • 한국생물공학회:학술대회논문집
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    • 2003.10a
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    • pp.810-814
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    • 2003
  • For the phylogenetic analysis of procaryotes, 16S rRNA gene has been used. In spite of it's common use, so high conservative of 16S rRNA gene limited resolving power, hence other molecule was applied in this study and the result was compared with that of 16S rRNA. COG (Clusters of Orthologous of protein) algorithm revealed that three COGs were only detected in 42 procaryotes ; transcription elongation factor (COG0195), bacterial DNA primase (COG0358) and uridylate kinase (COG0528). Uridylate kinase gene was selected owing to the similarity and one single copy number in each genome. Phylogenetic tree of 16S rRNA gene and uridylate kinase showed similarities and differences. Uridylate kinase may help the problem of very high conservative of 16S rRNA gene in rhylogenetic analysis and it would help to access more accurate discrimination and phylogenetic analysis of bacteria.

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Functional Analysis of PepRSH (Pepper relA/spoT homolog) cloned from Capsicum annuum showing Systemic Acquired Resistance against Phytophthora capsici

  • Kim, Tae-Ho;Kim, Yeong-Tae;Byun, Myung-Ok;Shin, Jeong-Sheop;Go, Seoung-Joo
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.69.1-69
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    • 2003
  • RSH (relA/spoT homolog) has been known to determine the level of guanosine tetraphosphate (ppGpp) and guanosine pentaphosphate (pppGpp), which are the effector nucleotide of the prokaryotic stringent response and also play a role in antibiotic production and differentiation in Streptomyces species but not a little in eukaryotic organism, especially in plant. Salicylic acid (SA), a critical signal molecule of establishing systemic acquired resistance (SAR), could induce SAR in Pepper (Capcicum annuum) against Phytophthora capsici. And the extent of SAR induction was in proportion to the dosage of SA (or BTH). Suppression subtractive hybridization (SSH), a PCR-based method for cDNA subtraction, was carried out between SA-treated and non-SA-treated pepper leaves to isolate genes which may be responsible for defense signaling against pathogens. Early upregulated gene was selected from reverse northern and kinetics of SSH-genes transcripts in SA-treated pepper leaves upon SA treatment. Full-length cDNA of the gene (PepRSH; Pepper RelA / SpoT homolog) had an open reading frame (ORF) of 2166 bp encoding a protein of 722 amino acids and a significant homology with (p)ppGpp phosphohydrolase or synthetase. Genomic DNA gel blot analysis showed that pepper genome has at least single copy of PepRSH. PepRSH transcripts was very low in untreated pepper leaves but strongly induced by SA and methyljasmonic acid (MeJA), indicating that PepRSH may share common SA and MeJA-mediated signal transduction pathway Functional analysis in E. coli showed PepRSH confers phenotypes associated with (p)ppGpp synthesis through a complementation using active site mutagenesis.

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STAT6 Gene Polymorphisms in Allergic Rhinitis

  • Kim, Jeong Joong;Kim, Min Su;Lee, Jung Hun;Choi, Tae Wook;Choi, Sang Heon;Lee, Jae Hoon;Chung, Hun Taeg
    • Genomics & Informatics
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    • v.2 no.3
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    • pp.126-130
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    • 2004
  • T helper-type 2 cytokines, such as IL-4 and IL-13, may play a central role in allergic diseases. The protein known as signal transducers and activators of transcription 6 (STAT6) is a key transcription factor involved in both IL-4- and -13-mediated biological responses. Two polymorphisms of the STAT 6 gene (exon 1 and G2964A variant) have been found. We investigated whether these STAT6 gene polymorph isms were associated with allergic rhinitis. Blood samples for genetic analysis were obtained from 285 individuals with allergic rhinitis and from 271 healthy subjects without atopic disease. The G2964A variant of the STAT6 gene was genotyped using PCR-RFLP analysis. The GT repeat polymorphism in exon 1 of the STAT6 gene was genotyped by fragment analysis. There was no association between the 2964A variant and GT repeat polymorphism in exon 1 of the STAT6 and allergic rhinitis in a Korean population (both p > 0.05). Our results suggest that a combination of STAT6 gene polymorphisms is not a useful marker for predicting allergic rhinitis.

Choristoneura fumiferana Granulovirus p74 Protein, a Highly Conserved Baculoviral Envelope Protein

  • Rashidan, Kianoush Khajeh;Nassoury, Nasha;Tazi, Samia;Giannopoulos, Paresa N.;Guertin, Claude
    • BMB Reports
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    • v.36 no.5
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    • pp.475-487
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    • 2003
  • A gene that encodes a homologue to baculoviral p74, an envelope-associated viral structural protein, has been identified and sequenced on the genome of Choristoneura fumiferana granulovirus (ChfuGV). A part of the ChfuGV p74 gene was located on an 8.9 kb BamHI subgenomic fragment using different sets of degenerated primers. These were designed using the results of the protein sequencing of a major 74 kDa structural protein that is associated with the occlusion-derived virus (ODV). The gene has a 1992 nucleotide (nt) open-reading frame (ORF) that encodes a protein with 663 amino acids with a predicted molecular mass of 74,812 Da. Comparative studies revealed the presence of two major conserved regions in the ChfuGV p74 protein. This study also shows that all of the p74 proteins contain two putative transmembrane domains at their C-terminal segments. At the nucleotide sequence level, two late promoter motifs (TAAG and GTAAG) were located upstream of the first ATG of the p74 gene. The gene contained a canonical poly(A) signal, AATAAA, at its 3' non-translated region. A phylogenetic tree for baculoviral p74 was constructed using a maximum parsimony analysis. The phylogenetic estimation demonstrated that ChfuGV p74 is related the closest to those of Cydia pomonella granulovirus (CpGV) and Phthorimaea operculella granulovirus (PhopGV).

A New Double-Stranded RNA Mycovirus from Pleurotus ostreatus (ASI 2504)

  • Lee, Jin-Kyung;Lee, Kang-Hyo;Shim, Hye-Kyung;Yang, Joo-Sung;Kim, Gyu-Hyun;Kong, Won-Sik;Yoo, Young-Bok;Kim, Dae-Hyun;Kim, Dong-Giun;Lee, Suk-Chan
    • The Plant Pathology Journal
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    • v.22 no.1
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    • pp.68-74
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    • 2006
  • A new virus with a dsRNA genome was isolated and characterized from the Suhan-:neutari strain (ASI 2504) of Pleurotus ostreatus, which was characterized as long and slightly bent with small caps on the stipe of fruit body. Thirty nm isometric viruses with three dsRNA segments (approximately 2.0, 1.84 and 1.82 kb in sizes) were isolated by ultracentrifugation in sucrose gradients. Western analysis of protein extracted purified viruses with anti-virus polyclonal antibody confirmed that viruses have two specific proteins (36 and 68 kDa). Computer analysis of 2.0 kb segment shows that high. sequence identity with RNA-dependent RNA polymerase (RdRp) of partitiviruses, respectively. When compared to other dsRNA mycoviruses in a phylogenetic analysis, OMDV was most related to Pleurotus ostreatus virus 1.

Risk assessment for clinical attachment loss of periodontal tissue in Korean adults

  • Rheu, Gun-Bak;Ji, Suk;Ryu, Jae-Jun;Lee, Jung-Bok;Shin, Chol;Lee, Jeong-Yol;Huh, Jung-Bo;Shin, Sang-Wan
    • The Journal of Advanced Prosthodontics
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    • v.3 no.1
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    • pp.25-32
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    • 2011
  • PURPOSE. The purpose of this study was to assess the prevalence and extent of clinical attachment loss of periodontal tissue and to find out variables related to clinical attachment loss (CAL) in Korean adults older than 40 years of age. MATERIALS AND METHODS. Data were collected from 2,519 subjects who were part of a cohort study conducted in Ansan city by Korea University Medical School for Korean Genome project. Age, sex, smoking, drinking, fast glucose, blood pressure, obesity and total cholesterol levels were examined. The oral examination included probing pocket depth, gingival recession and CAL of Ramford's teeth. The severity of periodontitis was classified based on the mean value of CAL. The relationship between each risk factor and the severity of CAL was independently estimated using the chi-square test, the test or one-way ANOVA. Multiple regression analysis was used to determine the significance of each factor in the periodontal disease. RESULTS. The prevalences of clinical attachment between 1 and 3 mm, between 3 and < 5 mm, and ${\geq}$ 5 mm were 80.27%, 16.75% and < 1%, respectively. Although the univariate analysis showed age, gender, smoking, fasting glucose, blood pressure and total cholesterol levels were significantly related to the severity of CAL, multiple regression analysis indicated that age (P < .0001), gender (P < .0001) and smoking (P < .05) were only significantly related. CONCLUSION. Older age, male gender and smoking were significant risk factor for the increase of CAL, and these may be useful indicators of periodontitis high-risk groups.

Phylogenetic Analysis of Apple scar skin viroid Isolates in Korea (국내에서 발생하는 Apple scar skin viroid 분리주에 대한 계통분석)

  • Cho, Kang Hee;Kim, In-Soo;Kil, Eui-Joon;Park, Seo Jun;Kim, Se Hee;Choi, In-Myung;Kim, Dae-Hyun
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.346-350
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    • 2015
  • To identify genome sequences of Apple scar skin viroid (ASSVd) isolates in Korea, the field survey was performed from 'Hongro' apple orchards located in eight sites in South Korea (Bongwha, Cheongsong, Dangjin, Gimchoen, Muju, Mungyeong, Suwon, and Yeongwol). ASSVd was detected by RT-PCR and PCR fragments were cloned into cloning vector. Full-length viral genomes of eight ASSVd isolates were sequenced and compared with 21 isolates reported previously from Korea, India, China, Japan and Greece. Eight isolates in this study showed 92.2-99.7% nucleotide sequence identities with those reported previously. Phylogenetic analysis showed that seven isolates reported in this study belong to the same group distinct from other groups.

Research Trend of Genetics in Oncology Nursing: Based on Text Network Analysis (유전종양간호 관련 연구경향: 텍스트 네트워크 분석을 중심으로)

  • Lee, Mijin;Oh, Soonyoung;Choi, Kyungsook
    • The Journal of the Korea Contents Association
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    • v.18 no.2
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    • pp.47-56
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    • 2018
  • The aim of this study is investigate the research trends by analyzing the researches related to Korean and international genetics in oncology nursing. We conducted a text network analysis focusing on the key words presented in the abstracts of papers published in journals related to genetics in oncology nursing. Nurse, Cancer, Genetic, Patient, Knowledge, Care, and Genetic Test were identified as keywords and centralized keywords. As a result of studying research trends over time, researches including keywords such as information, care, and knowledge have increased since the completion of the Human Genome Project in 2003. Key words classified through the meta paradigm of nursing were health, nursing, human, environment order. This study is meaningful in that it can be used to identify trends in tumor genetic nursing research and to set the direction of development of nursing intervention for hereditary cancer patients.