• Title/Summary/Keyword: gene-flow

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Silencing of the COPS3 Gene by siRNA Reduces Proliferation of Lung Cancer Cells Most Likely via induction of Cell Cycle Arrest and Apoptosis

  • Wang, Xue-Mei;Cui, Jiu-Wei;Li, Wei;Cai, Lu;Song, Wei;Wang, Guan-Jun
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.3
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    • pp.1043-1048
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    • 2012
  • The COPS3 gene has stimulating effect on cell proliferation and progression of osteosarcomas and related cells. However, the features of COPS3 and its potential application as a therapeutic target in other cancers has not yet been studied. In this study, therefore, the effect of COPS3 silencing via COPS3 siRNA on lung cancer cell proliferation was examined. Expression levels of COPS3 gene in COPS3 siRNA infected cells and control siRNA infected cells were compared with real time PCR and Western blot analysis. Cell proliferation levels were comprehensively analyzed by MTT, BrdU incorporationy, and colony formation assays. For mechanistic assessment the effects of COPS3 silencing on cell cycle and apoptosis were analyzed using flow cytometry. Results showed that successful silencing of the COPS3 gene at both translational and transcriptional levels significantly reduced the proliferation and colony formation by lung cancer cells (p<0.01). Flow cytometry showed cell cycle arrest in the G0/G1 phase after COPS3 silencing, and more importantly, apoptosis was induced as a result of COPS3 knockdown, which negatively affected cell survival. Therefore, these results provide another piece of important evidence that the COPS3 gene expressed in lung cancer cells may play a critical role in stimulating proliferation. Down-regulation of COPS3 could significantly inhibit lung cancer cell growth, which was most likely mediated via induction of cell cycle arrest in G0/G1 phase and apoptosis.

The Investigation of Gene Flows in Artificial Pollination between GM Rice and its Wild Relatives by RAPD Analysis (RAPD PCR에 의한 GM벼의 야생 근연종 벼로의 유전자 전이 분석법)

  • Kim, Yoon-Sik;Kim, Hyun-Soon;Joung, Hyouk;Jeon, Jae-Heung
    • Korean Journal of Plant Resources
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    • v.19 no.5
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    • pp.612-616
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    • 2006
  • In recent years, there has been increasing concerns in gene flow from GM crops to wild or weedy relatives as a potential risk in the commercialization of GM crops. To access the possibility of the environmental impacts by GM rice, small-scale experiments of gene transfer were carried out. Herbicide and drought stress resistant GM rice and non-GM rice Nakdongbyeo, wild rice Oryza nivara, and weedy rice Sharebyeo were used for artificial pollination experiments and bar gene was used as a tractable marker after pollination. The harvested putative hybrid seeds after artificial pollination were germinated and true hybrid plants were selected by basta treatment. The hybrid plants were verified again by PCR amplification of bar and trehalose-6-phosphate phosphatase (TPP) genes and RAPD PCR analysis.

The ybcF Gene of Escherichia coli Encodes a Local Orphan Enzyme, Catabolic Carbamate Kinase

  • Nam Yeun Kim;Ok Bin Kim
    • Journal of Microbiology and Biotechnology
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    • v.32 no.12
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    • pp.1527-1536
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    • 2022
  • Escherichia coli can use allantoin as its sole nitrogen source under anaerobic conditions. The ureidoglycolate produced by double release of ammonia from allantoin can flow into either the glyoxylate shunt or further catabolic transcarbamoylation. Although the former pathway is well studied, the genes of the latter (catabolic) pathway are not known. In the catabolic pathway, ureidoglycolate is finally converted to carbamoyl phosphate (CP) and oxamate, and then CP is dephosphorylated to carbamate by a catabolic carbamate kinase (CK), whereby ATP is formed. We identified the ybcF gene in a gene cluster containing fdrA-ylbE-ylbF-ybcF that is located downstream of the allDCE-operon. Reverse transcription PCR of total mRNA confirmed that the genes fdrA, ylbE, ylbF, and ybcF are co-transcribed. Deletion of ybcF caused only a slight increase in metabolic flow into the glyoxylate pathway, probably because CP was used to de novo synthesize pyrimidine and arginine. The activity of the catabolic CK was analyzed using purified YbcF protein. The Vmax is 1.82 U/mg YbcF for CP and 1.94 U/mg YbcF for ADP, and the KM value is 0.47 mM for CP and 0.43 mM for ADP. With these results, it was experimentally revealed that the ybcF gene of E. coli encodes catabolic CK, which completes anaerobic allantoin degradation through substrate-level phosphorylation. Therefore, we suggest renaming the ybcF gene as allK.

Genetic Variation in Korean Populations of Wild Radish, Raphanus sativus var.hortensis f. raphanistroides (Brassicaceae)

  • Hur, Man Kyu
    • Journal of Plant Biology
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    • v.38 no.4
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    • pp.329-336
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    • 1995
  • Raphanus sativus L. var. hortensis f. raphanistroides (wild radish: Brassicaceae), a herbaceous perennial, occurs only on beaches in East Asia. Genetic diversity and population structure of seven Korean populations were investigated using starch gel electrophoresis. Although the Korean populatins are small, isolated with patchy distribution, the population maintain a moderate level of genetic diversity; the mean percentage fo polymorphic loci was 51.4%, mean number of alleles per locus was 1.84, and mean expected heterozygosity was 0.116. A combination of animal-outcrossing breeding system, wide geographical distribution, restricted ecological distribution, and a propensity for high fecundity may in part be explanatory factors contributing the moderate level of genetic diversity within populations. An overall excess of homozygotes relative to Hardy-Weinberg expetations (mean FISa=0.116) indicates that consanguineous mating occur within wild radish populations, leading to a family structure within a circumscribed area. Although population of wild radish experience a limited gene flow, only 5% of the total genetic variation found in Korean wild radish populations examined is due to differences among populations (mean GST=0.052). This value is considerably lower than the mean values of species with similar life history and ecological characteristics. However, significant differences were found in allele frequencies between populations for all polymorphic loci (P<0.01). It is supposed that directional selection toward genetic uniformity (similar gene frequencies) in a relatively homogenous habitat is thought to be operated among Korean wild radish populations.

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Environmental risk assessment of genetically modified Herbicide-Tolerant zoysiagrass (Event: Jeju Green21) (제초제저항성 들잔디(Zoysia japonica Steud.) 이벤트 Jeju Green21의 환경위해성평가)

  • Bae, Tae-Woong;Kang, Hong-Gyu;Song, In-Ja;Sun, Hyeon-Jin;Ko, Suk-Min;Song, Pill-Soon;Lee, Hyo-Yeon
    • Journal of Plant Biotechnology
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    • v.38 no.2
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    • pp.105-116
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    • 2011
  • Transgenic zoysiagrass (Zoysia japonica Steud.) expressing the bar gene inserted in the plant genome has been generated previously through Agrobacterium tumefaciens-mediated transformation. The GM zoysiagrass (event: JG21) permits efficient management of weed control of widely cultivated zoysiagrass fields, reducing the frequency and cost of using various herbicides for weed control. Now we have carried out the environmental risk assessment of JG21 prior to applying to the governmental regulatory agency for the commercial release of the GM turf grass outside of test plots. The morphological phenotypes, molecular analysis, weediness and gene flow from each test plot of JG21 and wild-type zoysiagrasses have been evaluated by selectively analyzing environmental effects. There were no marked differences in morphological phenotypes between JG21 and wild-type grasses. The JG21 retained its stable integration in the host plant in T1 generation, exhibiting a 3:1 segregation ratio according to the Mendelian genetics. We confirmed the copy number (1) of JG21 by using Southern blot analysis, as the transgenic plants were tolerant to ammonium glufosinate throughout the culture period. From cross-fertilization and gene flow studies, we found a 9% cross-pollination rate at the center of JG21 field and 0% at distances over 3 m from the field. The JG21 and wild-type zoysiagrass plants are not considered "weed" because zoysiagrasses generally are not dominant and do not spread into weedy areas easily. We assessed the horizontal gene transfer (HGT) of the transgene DNA to soil microorganisms from JG21 and wild-type plants. The bar gene was not detected from the total genomic DNA extracted from each rhizosphere soil of GM and non-GM Zoysia grass fields. Through the monitoring of JG21 transgene's unintentional release into the environment, we found no evidence for either pollen mediated gene flow of zoysiagrass or seed dispersal from the test field within a 3 km radius of the natural habitat.

Genetic Structure in Korean Populations of Hosta capitata (Liliaceae)

  • Chung, Myong-Gi
    • Journal of Plant Biology
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    • v.37 no.3
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    • pp.277-284
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    • 1994
  • I investigated levels of genetic diversity, population genetic structure, and gene flow in Hostacapitata, a herbaceous perennial native to South Korea and southwestern Japan. Starch gel electrophoresis was conducted on leaves collected from 310 plants in 19 Korean populations. Twenty-two of 25 putative loci examined were polymorphic in at least one populatin and the mean number of alleles per locus was 1.65. In addition, mean expected heterozygosity within populations (Hep=0.153) was higher than average values for species with similar life history traits. Significant differences in allele frequency were detected between populations at all loci (P<0.01), and slightly over 30% of the genetic variation was found among populatins (GST=0.308). Indirect estimates of the number of migrants per generation (Nm) (0.506, calculated from GST; 0.852, calculated from the mean frequency of ten private alleles) indicate that gene flow is restricted among the isolated Korean populations of H. capitata. Factors contributing to the high levels of genetic differentiation among populations of H. capitata include small and discrete populations, human disturbance, and low frequencies of pollinator foraging behavior.

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Investigation of Possible Horizontal Gene Transfer from Transgenic Rice to Soil Microorganisms in Paddy Rice Field

  • Kim, Sung-Eun;Moon, Jae-Sun;Kim, Jung-Kyu;Choi, Won-Sik;Lee, Sang-Han;Kim, Sung-Uk
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.187-192
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    • 2010
  • In order to monitor the possibility of horizontal gene transfer between transgenic rice and microorganisms in a paddy rice field, the gene flow from a bifunctional fusion (TPSP) rice containing trehalose-6-phosphate synthase and phosphatase to microorganisms in soils was investigated. The soil samples collected from the paddy rice field during June 2004 to March 2006 were investigated by multiplex PCR, Southern hybridization, and amplified fragment length polymorphism (AFLP). The TPSP gene from soil genomic DNAs was not detected by PCR. Soil genomic DNAs did not show homologies on the Southern blotting data, indicating that gene transfer did not occur during the last two years in the paddy rice field. In addition, the AFLP band patterns produced by soil genomic DNAs from both transgenic and non-transgenic rice fields appeared similar to each other when analyzed by the NTSYSpc program. Thus, these data suggest that transgenic rice does not give a significant impact on the communities of soil microorganisms, although long-term observation may be needed.

Role of Kupffer Cells in Alteration of Vasoregulatory Gene Expression in Hepatic Ischemia/Reperfusion

  • Kim, Yong-Hyuk;Kim, Sung-Ho;Lee, Sun-Mee
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.299.1-299.1
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    • 2002
  • Failure of the hepatic microcirculation is a major component of reperfusion injury in the liver. However. the vasoactive mediators involved in the regulation of sinusoidal flow during reperfusion following hepatic ischemia remain to be identified. We investigate the role of Kupffer cells in hepatic ischemia/reperfusion (l/R)-induced imbalance of vasoregulatory gene expression. Rats were subjected to 60 min hepatic ischemia, followed by 5 h of reperfusion. (omitted)

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Genetic Diversify and Population Structure of Two Korean Pond Frog Species, Rana nigromaculata and R. plancyi (Anura, Ranidae), with a Survey of Temporal Genetic Variation in R. nigromaculata

  • Suh-Yung Yang;Jong-Bum Kim;Mi-Sook Min;Jae-Hwa Suh
    • Animal cells and systems
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    • v.3 no.3
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    • pp.275-283
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    • 1999
  • Korean R. plancyi occupies a restricted area in western South Korea and shows a relatively low level of genic variability (%P=15.2, Ho=0.052, He=0.048). In contrast, R. nigromaculata is broadly distributed in South Korea. The observed low level of variability of R. nigromaculata (%P=14.3, Ho=0.042, He=0.043) is probably due to its recent colonization. Populations of R. nigromaculata exhibited considerable genetic differentiation (F$_{sT}$=0.149) and low level of gene flow (Nm=1.427) among populations, compared to those of R. Plancyi (F$_{sTF$_{sT}$}$=0.096, Nm=2.354), which occupies a restricted area. The observed levels of gene flow among populations of R. nigromaculata (Nm=1.427) over a broad geographic range is relatively higher than other amphibian species. The high level of gene flow is probably the result of the high dispersal abilities of R. nigromaculata. A survey of temporal genic variation of R. nigromaculata showed that there was no significant change on the overall average genetic diversity from 1978 (average He=0.044) to 1997 (average He=0.040). Wright's F-statistics also indicated no significant genetic differentiation from 1978 (F$_{sT}$=0.118) to 1997 (F$_{sT}$=0.108). This suggests that the environmental change appears to have had little influence on the genetic composition of R. nigromaculata in the study areas during the past 20 years. The low level of temporal variation might be due to the result of high dispersal abilities and wide migration range of this species.

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Influence of gene flow from GM to non-GM soybeans by the size of the pollen donor

  • Lee, Bumkyu;Oh, Sung-Dug;Chang, Ancheol
    • Korean Journal of Agricultural Science
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    • v.45 no.4
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    • pp.591-600
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    • 2018
  • The use of genetically modified (GM) crops has increased continuously over the world, and concerns about the potential risks of GM crops have also risen. Although, until now, GM crops have not been cultivated commercially in Korea, it is necessary to develop technology for the safe evaluation of GM crops. In this study, we investigated the influence of gene flow from GM to non-GM soybeans by the size of the pollen donor. In the experimental design, GM soybeans were placed in the center as a pollen donor and non-GM soybeans were placed in four directions as the pollen receivers. Three sizes of pollen donor were designed as $90cm{\times}90cm$, $180cm{\times}180cm$, and $360cm{\times}360cm$. A total 22,719 seeds were collected from non-GM soybeans, and 14 hybrids were finally obtained through herbicide resistance screening and PCR analysis. The highest hybridization rate was 0.78% at a distance of 15 cm from a $360cm{\times}360cm$ GM pollen donor, and the farthest distance of hybridization was 180 cm from a GM pollen donor which was $360cm{\times}360cm$ in size. Ten hybrids were found among the 14 hybrids at the $360cm{\times}360cm$ pollen donor size, 3 hybrids at $180cm{\times}180cm$, 1 hybrid at $90cm{\times}90cm$. From these results, it could be concluded that with the larger pollen donor size, more hybridization occurred in soybeans.