• Title/Summary/Keyword: Gene Screening

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Cytoprotection Against Oxidative Damage by Nrf2-regulated Genes

  • Kwak, Mi-Kyoung;Kensler, Thomas W.
    • Toxicological Research
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    • v.23 no.3
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    • pp.207-214
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    • 2007
  • Chronic oxidative stress produced by exposure to environmental chemicals or pathophysiological states can lead animals to aging, carcinogenesis and degenerative diseases. Indirect antioxidative mechanisms, in which natural or synthetic agents are used to coordinately induce the expression of cellular antioxidant capacity, have been shown to protect cells and organisms from oxidative damages. Electrophile and free radical detoxifying enzymes, which were originally identified as the products of genes induced by cancer chemopreventive agents, are members of this protective system. The NFE2 family transcription factor Nrf2 was found to govern expression of these detoxifying enzymes, and screening for Nrf2-regulated genes has identified many gene categories involved in maintaining cellular redox potential and protection from oxidative damage as Nrf2 downstream genes. Further, studies using Nrf2-deficient mice revealed that these mutant mice showed more susceptible phenotypes towards exposure to environmental chemicals/carcinogens and in oxidative stress related disease models. With the finding that cancer chemopreventive efficacy of indirect antioxidants (enzyme inducers) is lost in the absence of Nrf2, a central role of Nrf2 in the antioxidative protective system has been firmly established. Promising results from cancer prevention clinical trials using enzyme inducers propose that pharmacological interventions that modulate Nrf2 can be an effective strategy to protect tissues from oxidative damage.

Phylogenetic Analysis of New Isolates of Cucumber mosaic virus from Iran on the Basis of Different Genomic Regions

  • Nematollahi, Sevil;Sokhandan-Bashir, Nemat;Rakhshandehroo, Farshad;Zamanizadeh, Hamid Reza
    • The Plant Pathology Journal
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    • v.28 no.4
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    • pp.381-389
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    • 2012
  • Molecular characterization of Cucumber mosaic virus (CMV) was done by using samples from tomato and cucurbitaceous plants collected from different locations in the northwest region of Iran. After screening by enzyme-linked immunosorbent assay, 91 CMV-infected samples were identified. Biological properties of eight representative isolates were compared with each other revealing two distinct phenotypes on squash and tomato plants. Phylogenetic analyses based on nucleotide sequences of the coat protein (CP), movement protein (MP) and 2b of the new isolates, together with that of previously reported isolates, led to the placement of the Iranian isolates in subgroups IA and IB according to CP and MP genes, but in subgroup IA according to the 2b gene. These data suggest that reassortment may have been a major event in the evolution of CMV in Iran, and that the Iranian isolates are derived from a common recent ancestor that had passed through a bottleneck event.

Isolation, Identification, and Characterization of Bacillus strains from the Traditional Korean Soybean-fermented Food, Chungkookjang

  • Joo, Myeong-Hoon;Hur, Sung-Ho;Han, Yong-Soo;Kim, Ji-Yeon
    • Journal of Applied Biological Chemistry
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    • v.50 no.4
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    • pp.202-210
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    • 2007
  • A total of 45 bacterial strains were isolated from the traditional Korean soybean-fermented food, Chungkookjang. Among these strains, seven strains were selected and identified based on morphological, physiological, and biochemical characteristics, as well as phylogenetic analysis using 16S rDNA sequences. All strains were Gram-positive, aerobic, motile, oxidase-positive, rod-shaped, and endospore-forming bacteria, and produced extracellular enzymes such as amylase, cellulase, lipase, protease, and xylanase. The isolates were grown in the presence of 0-11% (w/v) NaCl. Growth was optimal at pH 6-9 and at temperatures of $30-45^{\circ}C$. According to VITEK automicrobic system tests and supplementary tests, the isolates were similar to several species of the genus Bacillus. The phylogenetic analysis of seven bacterial strains based on comparisons of 16S rDNA sequences, revealed that the strains were closely related to Bacillus species. The identification of strains that produced surfactin was also carried out, based on PCR screening of the sfp gene. Among the seven isolated strains, six yielded a surfactin-positive result with PCR.

Discrimination of Hantaviruses from the Tissues of Infected Hamsters to 5 Different Serotype Hantaviruses by Nested RT-PCR using Hantavirus Serotype Specific Primers (한타바이러스 혈청형 특이 Primer를 이용한 Nested RT-PCR 방법으로 5가지 혈청형 한타바이러스에 감염된 햄스터 조직에서 바이러스 검출)

  • Chu, Yong-Kyu;Lee, Ho-Wang
    • The Journal of Korean Society of Virology
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    • v.27 no.1
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    • pp.49-57
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    • 1997
  • We developed a sensitive, nested reverse transcription-polymerase chain reaction (RT-PCR) to detect Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in animal tissues. Total RNA was extracted from blood, lung or kidney samples of experimentally-infected hamsters by using the guanidine isothiocyanate buffer-acid phenol-chloroform method. Genus-reactive outer primers were derived from the consensus region of the G1 gene sequences of several hantaviruses. Serotype-specific primers were selected within the region amplified by the outer primers. To examine the sensitivity and specificity of the test, we diluted known quantities of Hantaan, Seoul, Belgrade, Puumala and Sin Nombre viruses in human or hamster immune sera before performing the nested RT-PCR. We could detect as little as 1 pfu of virus, even in the presence of high-titer neutralizing antibodies, and the serotype-specific primers amplified only homologous serotype viruses. RT-PCR with these primers demonstrated virus in the blood of experimentally-infected hamsters as early as four days to as late as 30 days after infection. A comparison of a standard immunofluorescent antibody screening test (IFAT) to nested RT-PCR with RNA extracted from lung or kidney tissues of the hamsters, demonstrated that RT-PCR to be more sensitive for identifying viruses in these tissues.

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Characterization of Diesel Degrading Enterobacter cancerogenus DA1 from Contaminated Soil

  • Kim, Sang-Jun;Joo, Gil-Jae
    • Korean Journal of Environmental Biology
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    • v.36 no.2
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    • pp.190-198
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    • 2018
  • The petroleum industry is an important part of the world economy. However, the massive exposure of petroleum in nature is a major cause of environmental pollution. Therefore, the microbial mediated biodegradation of petroleum residues is an emerging scientific approach used to resolve these problem. Through the screening of diesel contaminated soil we isolated a rapid phenanthrene and a diesel degrading bacterium identified as Enterobacter cancerogenus DA1 strain through 16S rRNA gene sequence analysis. The strain was registered in NCBI with an accession number MG270576. The optimal growth condition of the DA1 strain was determined at pH 8 and $35^{\circ}C$, and the highest degradation rate of the diesel was achieved at this condition. At the optimal condition, growth of the strain on the medium containing 0.05% phenanthrene and 0.1% of diesel-fuel was highest at 45 h and 60 h respectively after the incubation period. Biofilm formation was found significantly higher at $35^{\circ}C$ as compared to $30^{\circ}C$ and $40^{\circ}C$. Likewise, the lipase activity was found significantly higher at 48 h after the incubation compared to 24 h and 72 h. These results suggest that the Enterobacter cancerogenus DA1 could be an efficient candidate, for application through ecofriendly scientific approach, for the biodegradation of petroleum products like diesel.

Isolation and Identification of Lactic Acid Bacteria from Sourdough with High Exopolysaccharide Production Ability

  • Jung, Seung-Won;Kim, Wang-June;Lee, Kwang-Geun;Kim, Cheol-Woo;Noh, Wan-Seob
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.384-389
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    • 2009
  • To isolate lactic acid bacteria having high exopolysaccharides (EPS) production ability, 50 strains were initially isolated from the sourdough. Twenty-one strains formed highly mucoid colonies on the sucrose agar medium, which are indicative of active EPS synthesis. DU-07, DU-10, DU-12, DU-19, and DU-21 produced $11.51{\pm}0.167$, $13.09{\pm}0.193$, $12.72{\pm}0.108$, $11.61{\pm}0.284$, and $13.32{\pm}0.094\;g/L$ EPS, respectively, in MRS medium. The isolated strains, DU-10, DU-12, and DU-21, were identified as Enterococcus flavescens, Enterococcus faecium, and Lactobacillus amylovorus, respectively, by using API 50CHL kit and determining partial sequences of their 16S rDNA. Especially, L. amylovorus DU-21 showed the highest production of EPS, as well as the highest inhibitory activities against pathogenic (p<0.05). Interestingly, the L. amylovorus DU-21 seem to be endemic to sourdough fermentations, as they have not been isolated from other environments.

Genetic testing in clinical pediatric practice

  • Yoo, Han Wook
    • Clinical and Experimental Pediatrics
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    • v.53 no.3
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    • pp.273-285
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    • 2010
  • Completion of the human genome project has allowed a deeper understanding of molecular pathophysiology and has provided invaluable genomic information for the diagnosis of genetic disorders. Advent of new technologies has lead to an explosion in genetic testing. However, this overwhelming stream of genetic information often misleads physicians and patients into a misguided faith in the power of genetic testing. Moreover, genetic testing raises a number of ethical, legal, and social issues. Diagnostic genetic tests can be divided into three primary but overlapping categories: cytogenetic studies (including routine karyotyping, high-resolution karyotyping, and fluorescent in situ hybridization studies), biochemical tests, and DNA-based diagnostic tests. DNA-based testing has grown rapidly over the past decade and includes preandpostnatal testing for the diagnosis of genetic diseases, testing for carriers of genetic diseases, genetic testing for susceptibility to common non-genetic diseases, and screening for common genetic diseases in a particular population. Theoretically, once a gene's structure, function, and association with a disease are well established, the clinical application of genetic testing should be feasible. However, for routine applications in a clinical setting, such tests must satisfy a number of criteria. These criteria include an acceptable degree of clinical and analytical validity, support of a quality assurance program, possibility of modifying the course of the diagnosed disease with treatment, inclusion of pre-and postnatal genetic counseling, and determination of whether the proposed test satisfies cost-benefit criteria and should replace or complement traditional tests. In the near future, the application of genetic testing to common diseases is expected to expand and will likely be extended to include individual pharmacogenetic assessments.

Screening of Yeast Diauxic Promoters for Production of Foreign Proteins

  • Kim Jin-Ju;Kim Sang-Woo;Jeon Che-Ok;Yun Ji-Yun;Lee Hyun-Sook;Ro Hyeon-Su
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1459-1463
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    • 2006
  • This study explored yeast diauxic promoters using a green fluorescent protein (GFP) reporter to screen growth phase-controlled promoters applicable for foreign protein production. Twenty-five diauxic promoters were inserted into a yeast 2-micron vector in front of the reporter GFP gene. The expressed GFP signal intensity measurements showed that 23 out of the 25 promoters produced a significant fluorescent signal when the cells were in the diauxic growth phase. Among the two strongest promoters pYDL204W and pYLR258W, the former remained constantly active after its activation at the diauxic shift, whereas the latter was only transiently activated right after the deprivation of the medium glucose.

Cloning and Characterization of a Novel ${\alpha}$-Amylase from a Fecal Microbial Metagenome

  • Xu, Bo;Yang, Fuya;Xiong, Caiyun;Li, Junjun;Tang, Xianghua;Zhou, Junpei;Xie, Zhenrong;Ding, Junmei;Yang, Yunjuan;Huang, Zunxi
    • Journal of Microbiology and Biotechnology
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    • v.24 no.4
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    • pp.447-452
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    • 2014
  • To isolate novel and useful microbial enzymes from uncultured gastrointestinal microorganisms, a fecal microbial metagenomic library of the pygmy loris was constructed. The library was screened for amylolytic activity, and 8 of 50,000 recombinant clones showed amylolytic activity. Subcloning and sequence analysis of a positive clone led to the identification a novel gene (amyPL) coding for ${\alpha}$-amylase. AmyPL was expressed in Escherichia coli BL21 (DE3) and the purified AmyPL was enzymatically characterized. This study is the first to report the molecular and biochemical characterization of a novel ${\alpha}$-amylase from a gastrointestinal metagenomic library.

Healing of Bony Defects by Cervi Pantotricuhum Cornu Herbal Acupuncture (녹용약침(鹿茸藥鍼)이 골형성(骨形成)에 미치는 영향(影響))

  • Han, Sang-Won;Choi, Je-Yong;Lee, Yun-Ho
    • Journal of Acupuncture Research
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    • v.18 no.5
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    • pp.135-146
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    • 2001
  • Objective : Bone homeostasis is maintained by balance of bone formation and resorption. Therefore, bone related diseases arose by disturbance of this balance between osteoblast and osteoclast activities. To develop a successful screening system of the therapeutic components based on oriental medicine is essential to set out systematic approach for that purpose. Methods : This study was perforated Sprague-Dawley rat femur for bony defects(${\phi}5mm$) by the fissure bur. And experimetal group were treated with Cervi Pantotricuhum Cornu injection at both $Sh{\grave{e}}nsh{\tilde{u}}$(BL23) & $D{\grave{a}}zh{\grave{u}}(BL11)(0.2m{\ell})$. This was evaluated by radiography and histological analysis with in situ hybridization. Results : Cervi Pantotricuhum Cornu Herbal Acupuncture has weak effect on bony defect healing and this was evaluated by X-ray taking and histological analysis with in situ hybridization. Osteocalcin gene expression was not changed by Cervi Pantotricuhum Cornu Herbal Acupuncture in bony defects animal model. Conclusion : Taken together this study show that the Cervi Pantotricuhum Cornu herbal-acupuncture has a weak effect healing of bony defects, this type of approach might give a good chance to explore the favorable effects of Cervi Pantotricuhum Cornu herbal-acupuncture on bone tissue.

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