• Title/Summary/Keyword: mechanism for gene flow

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Mediation of Gene Flow in Tropical Trees of Sub-Saharan Africa

  • Onokpise, Oghenekome U.;Akinyele, Adejoke O.
    • Journal of Forest and Environmental Science
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    • v.28 no.1
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    • pp.1-7
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    • 2012
  • Tropical forests whether fragmented or undisturbed or be they equatorial or deciduous, remain the storehouse of biodiversity for hundreds of thousands of plant and animal species. This unique characteristic continues to attract a wide range of scientists and international organizations to study and attempt to understand tropical forest ecosystems. Gene flow is mediated by pollen, seed and seedling dispersal, and factors affecting this gene flow include phenology, spatial distribution, population structures, seed predation, sexual and mating systems as well as physical and biological barriers to gene flow. Two methods are used in measuring gene flow: direct method that relies on the actual observation of seed and pollen dispersal, whereas indirect methods involve the use of genetic markers such as allozymes and DNA techniques. Political strife, extreme natural and artificial disasters, the lack of a comprehensive forestry research vision, coupled with difficult socio-economic conditions in Africa have made the environment quite difficult for sustained research activities on the part of those undertaking or wishing to undertake such studies. Gene flow studies in this region are few and far between. This review elaborates on the mechanisms of gene flow mediation in Sub-Saharan Africa.

Utilization of the bar gene to develop an efficient method for detection of the pollen-mediated gene flow in Chinese cabbage (Brassica rapa spp. pekinensis)

  • Lim, Chaewan;Kim, Sunggil;Choi, Yeonok;Park, Young-doo;Kim, Sung Uk;Sung, Soon-Kee
    • Plant Biotechnology Reports
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    • v.1 no.1
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    • pp.19-25
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    • 2007
  • To develop an efficient screening method for detection of the transgene in Chinese cabbage (Brassica rapa spp. pekinensis) utilizing Basta spray, optimal conditions for Basta application were examined in this study. Two transgenic Chinese cabbage lines were obtained through Agrobacterium-mediated transformation and used as transgenic positive controls in the Basta screening experiment. Differential concentrations of glufosinate-ammonium were sprayed into three different growth stages of 12 commercial Chinese cabbage cultivars. The results showed that no plants could survive higher than 0.05% glufosinate-ammonium, and plants at the 2-3 leaf stage were most vulnerable to glufosinate-ammonium. On the other hand, no damage was observed in the transgenic control plants. Reliability of the Basta spray method was proven by showing perfect co-segregation of the tolerance to glufosinate-ammonium and the presence of the bar gene in T1 segregating populations of the transgenic lines, as revealed by both PCR and Southern blot analyses. Using the developed Basta screening method, we tried to investigate the transgene flow through pollen dispersal, but failed to detect any transgene-containing non-transgenic Chinese cabbages whose parents had been planted adjacent to transgenic Chinese cabbages in field conditions. However, the transgene was successfully detected using Basta spray from the non-transgenic plants bearing the transgene introduced by hand-pollination. Since the Basta spray method developed in this study is easy to apply and economical, it will be a valuable tool for understanding the mechanism of gene flow through pollen transfer and for establishing a biosafety test protocol for genetically modified (GM) Chinese cabbage cultivars.

Genetic Structure of Macrophomina phaseolina Populations, the Causal Agent of Sesame Charcoal Rot Disease in Iran

  • Maryam Dolatkhah;Fariba ghaderi;Abdollah Ahmadpour
    • Research in Plant Disease
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    • v.30 no.1
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    • pp.50-59
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    • 2024
  • Charcoal rot disease, caused by the fungus Macrophomina phaseolina, is one of the most important diseases of Sesame (Sesamum indicum) all over the world. However, the population biology of M. phaseolina is poorly understood. In this study, M. phaseolina isolates from five different regions of Iran (Khuzestan, Fars, Bushehr, Hormozgan, and Kohgiluyeh & Boyer-Ahmad provinces) (n=200) were analyzed for genetic variation using inter simple sequence repeats marker. In total, 152 unique haplotypes were identified among the 200 M. phaseolina isolates, and gene diversity (H=0.46-0.84) and genotypic diversity were high in each of the regions. The structure analysis clustered five Iranian populations into two distinct groups, the individuals from group 1 were assigned to the Bushehr population and the individuals from Khuzestan, Fars, Hormozgan and Kohgiluyeh & Boyer-Ahmad were aggregated and formed group 2. The results matched with genetic differentiation and gene flow among regions. Analyses of the distribution of gene diversity within and among five Iranian populations were 61% and 39%, respectively. Our results showed that infected seeds are thought to be the dominant mechanism responsible for the spreading of the pathogen in southern parts of Iran. In summary, it is essential to have local quarantine and prevent seed exchanges between geographical populations to restrict the dispersal of pathogen over long distances and provide certified seeds in Iran.

Investigation of Possible Horizontal Gene Transfer from the Leaf Tissue of Transgenic Potato to Soil Bacteria

  • KIM YOUNG TAE;KIM SUNG EUN;PARK KI DUK;KANG TAE HOON;LEE YUN MI;LEE SANG HAN;MOON JAE SUN;KIM SUNG UK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1130-1134
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    • 2005
  • To monitor the possibility of horizontal gene transfer between transgenic potato and bacteria in the environment, the gene flow from glufosinate-tolerant potato to bacteria in soils was investigated. The soil samples treated with the leaf tissue of either glufosinate-tolerant or glufosinate-sensitive potato were subjected to PCR and Southern hybridization to determine possible occurrence of glufosinate-resistant soil bacteria and to detect the bar (phosphinothricin acetyltransferase) gene, conferring tolerance to glufosinate. The bar gene was not detected from genomic DNAs extracted at different time intervals from the soil samples, which had been treated with the leaf tissue of either transgenic or non-transgenic potato for 2 to 8 weeks. In addition, the level of glufosinate-resistant bacteria isolated from the soil samples treated with the leaf tissue of transgenic potato was similar to that of the samples treated with non-transgenic potato after 4 months of incubation at $25^{\circ}C$. The bar gene was not detected in the genomic DNAs extracted from colonies growing on the plate containing glufosinate, indicating that the bacteria could acquire the resistant phenotype to glufosinate by another mechanism without the uptake of the bar gene from glufosinate-tolerant potato.

Identification of Differentially Expressed Genes (DEGs) by Malachite Green in HepG2 Cells

  • Kim, Youn-Jung;Song, Mee;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.4 no.1
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    • pp.22-30
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    • 2008
  • Malachite Green (MG), a toxic chemical used as a dye, topical antiseptic and antifungal agent for fish, is highly soluble in water, cytotoxic to various mammalian cells and also acts as a liver tumor promoter. In view of its industrial importance and possible exposure to human beings, MG possesses a potential environmental health hazard. So, we performed with HepG2, a human hepatocellular carcinoma cell line, to identify the differentially expressed genes (DEGs) related to toxicity of MG. And we compared gene expression between control and MG treatment to identify genes that are specifically or predominantly expressed by employing annealing control primer (ACP)-based $GeneFishing^{TM}$ method. The cytotoxicity $(IC_{20})$ of MG was determined above the $0.867{\mu}M$ in HepG2 cell for 48 h treatment. And the DEGs of MG were identified that 5 out of 6 DEGs were upregulated and 1 out of 6 DEGs was down-regulated by MG. Also, MG induced late apoptosis and necrosis in a dose dependent in flow cytometric analysis. Through further investigation, we will identify more meaningful and useful DEGs on MG, and then can get the information on mechanism and pathway associated with toxicity of MG.

The effect of Leonurus sibiricus on the proliferation inhibition of human uterine leiomyoma cell and expression of gene related cell apoptosis (익모초(益母草)가 자궁근종세포의 증식억제와 Apoptosis 관련 유전자 발현에 미치는 영향)

  • Lee, Soo-Jin;Beak, Seung-Hee;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.3
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    • pp.1-12
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    • 2006
  • Purpose : This study was aimed to investigate the inhibitory effect of Leonurus sibiricus on the proliferation of human uterine leiomyoma cells and the expression of gene related the mechanism of cell apoptosis. Methods : We counted the number of death cells treated with indicated concentration of Leonurus sibiricus and investigated cell death rate by MTS assay. Furthermore, flow cytometry analysis and DNA fragmentation assay were used to dissect between necrosis and apoptosis and then we observed the differential gene expression by western blot analysis. Results : Leonurus sibiricus significantly inhibited the proliferation of uterine leiomyoma cell in a dose-dependent and time dependent manner. Fluorescence activated cell sorter (FACS) analysis indicated that Leonurus sibiricus induced G1 cell cycle arrest. Leonurus sibiricus enhanced the expression of p27 and p53 with cell cycle arrest. Conclusion : These findings suggest that Leonurus sibiricus is a candidate agent for the treatment of uterine leiomyoma. p27, $p53^{1}$ may play an important role in Leonurus sibiricus-induced cell cycle arrest and cell growth inhibition.

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Effect of Hyulbuchukeotang on the Inhibition of Proliferation of Uterine leiomyoma cells and Cell apoptosis (혈부축어탕(血府逐瘀湯) 자궁근종세포의 증식억제와 Apoptosis 관련 유전자 발현에 미치는 영향)

  • Moon, Na-Young;Baek, Seung-Hee;Kim, Dong-Chul
    • The Journal of Korean Obstetrics and Gynecology
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    • v.19 no.2
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    • pp.186-198
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    • 2006
  • Purpose : The purpose of this study is to demonstrate the direct inhibitory effect of Hyulbuchukeotang on the proliferation of uterine leiomyoma cells through an experiment treating uterine leiomyoma cells cultivated by explantation with indicated concentrations of Hyulbuchukeotang and to research the gene expression related to cell cycle ill order to discover the connection with apoptosis and its mechanism by analyzing cell cycle. Methods : After primary culture of uterine leiomyoma cells, the cultivated uterine leiomyoma cells were treated with indicated concentrations of Hyulbuchukeotang for 24 hours. The inhibitory effect on the cell proliferation was determined by the cell count assay. The value of a cell count assay represent the percentage of cells in a phase of the cell cycle compared with total cells. In addition, a link between Hyulbuchukeotang and apoptosis was examined through flow cytometric analysis by FACS and DNA fragmentation analysis. Finally, the degree of gene expression related to cell cycle was evaluated by Western blot analysis. Results : The inhibitory effect of Hyulbuchukeotang increase of uterine leiomyoma cells treated with indicated concentrations of Hyulbuchkeotang increases. The result of gene expression related to G1 phase after treating with 100, 250, 500, 1,000 ${\mu}g/ml$ concentrations of Hyulbuchukeotang. on uterine leiomyoma cells is that the gene expression of p27 was increased but that of p53 an p21 remained unchanged and the gene of pRB, pro-caspase 3 was decreased. Conclusion Through the mentioned experiments, it is demonstrated that Hyulbuchkeotang is effective in inhibiting Proliferation of uterine leiomyoma cells by extending cell cycle G1. However it is not considered that the inhibitory effect results from the aptoposis.

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Molecular Biological Study of Anti-cancer Effects of Bee Venom Aqua-acupuncture (봉독약침(蜂毒藥鍼)의 항암효과(抗癌效果)에 대한 분자생물학적(分子生物學的) 연구(硏究))

  • Park, Chan-Yol;Seo, Jung-Chul;Choi, Do-Young;Ahn, Byoung-Choul
    • Journal of Pharmacopuncture
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    • v.3 no.1
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    • pp.1-19
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    • 2000
  • To study anti-cancer effect and molecular biological mechanism of bee venom for aqua-acupuncture, the effects of bee venom on cell viability and apoptosis were analyzed using MTT assay, tryphan blue assay, $[^3H]$thymidine release assay, flow cytometric analysis, and activity of caspase-3 protease activity assay. To explore whether anti-cancer effects of bee venom are associated with the transcriptional control of gene expression, quantitative RT-PCR analysis of apoptosis-related genes was performed. The obtained results are summarized as follows: 1. The MTT assay demonstrated that cell viability was decreased by bee venom in a dose-dependant manner. 2. Significant induction of apoptosis was identified using tryphan blue assay, $[^3H]$thymidine release assay, and flow cytomet1 ric analysis of sub $G_1$ fraction. 3. In analysis of caspase-3 protease activity, the activity had increased significantly, in a dose-dependant manner. 4. Quantitative RT-PCR analysis of the apoptosis-related genes showed that Bcl-2 and Bcl-$X_L$ were down-regulated whereas Bax was up-regulated by bee venom treatment.

Immunity Responses of the Spatholubus Suberectus Dunn to the Synovial Cells Isolated from Patients with Rheumatoid Arthritis (류마토이드 관절염 환자 골막세포에 대한 계혈등의 면역반응)

  • Seo Hae Gyoung;Oh Min Suck;Kim Dong Hee
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.3
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    • pp.780-786
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    • 2003
  • This study was carried out to know the immunity responses of the Spatholubus suberectus Dunn(hereinafter referred to STSD) to the synovial cells isolated from patients with rheumatoid arthritis. Various experiments were performed in vitro to analyse the immunity effects of STSD. Gene expression and production of pro-inflammatory cytokines such as IL-1β, IL-6, TNF-α, iNOS and COX-2 were determined by RT-PCR and ELISA kit. And also the binding activity of NF-kB and AP-1 were measured by Electromobility shift assay (EMSA) and the production of ROS was measured by flow cytometry. The results were obtained as follows 1. The gene expression and production of pro-inflammatory cytokines IL-1β, IL-6, TNF-α were reduced significantly. 2. The gene expression of iNOS and COX-2 were reduced. 3. The binding activity of NF-kB and AP-1 were inhibited. 4. The production of ROS in human synovial cells was reduced significantly. Comparison of the results for this study showed that STSD had immunomodulatory effects of suppressing or enhancing. So we expect that STSD should be used as a effective drugs for not only rheumatoid arthritis but also another auto-immune disease. Therefore we have to survey continuously in looking for the effective substance and mechanism in the future.

Up-Regulation of Interleukin-4 Receptor Expression by Interleukin-4 and CD40 Ligation via Tyrosine Kinase-Dependent Pathway

  • Kim, Hyun-Il;So, Eui-Young;Yoon, Suk-Ran;Han, Mi-Young;Lee, Choong-Eun
    • BMB Reports
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    • v.31 no.1
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    • pp.83-88
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    • 1998
  • Recently a B cell surface molecule, CD40, has emerged as a receptor mediating a co-stimulatory signal for B cell proliferation and differentiation. To investigate the mechanism of synergy between interleukin-4 (IL-4) and CD40 ligation in B cell activation, we have examined the effect of CE40 cross-linking on the IL-4 receptor expression in human B cells using anti-CE40 antibody. We observed that IL-4 and anti-CD40 both induce IL-4 receptor gene expression with a rapid kinetics resulting in a noticeable accumulation of IL-4 receptor mRNA within 4 h. While IL-4 caused a dose-dependent induction of surface IL-4 receptor expression, the inclusion of anti-CD40 in the IL-4-treated culture, further up-regulated the IL-4-induced IL-4 receptor expression as analyzed by flow cytometry. Pretreatment of B cells with inhibitors of protein tyrosine kinase (PTK) resulted in a significant inhibition of both the IL-4- and anti-CD40-induced IL-4 receptor mRNA levels, while protein kinase C (PKC) inhibitors had no effects. These results suggest that IL-4 and CD40 ligation generate B cell signals, which via PTK-dependent pathways, lead to the synergistic induction of IL-4 receptor gene expression. The rapid induction of IL-4 receptor gene expression through the tyrosine kinase-mediated signal transduction by B cell activating stimuli, would provide cells capacity for an efficient response to IL-4 in the early phase of IL-4 action, and may in part constitute the molecular basis of the reported anti-CD40 co-stimulatory effect on the IL-4-induced response.

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