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DD-PCR응 이용한 벤조피렌 노출 조피볼락의 차등발현 유전자 분석
염승식,우선옥,최은석,김소정,오로라,이석찬,이택견 환경독성보건학회 2005 환경독성보건학회지 Vol.20 No.1
오염물질의 노출에 의해 발현이 변화되는 유전자의 발굴은 외부환경 자극에 대한 적응이나 반응의 메커니즘을 알아내는데 중요한 정보를 제공하며, 오염물질에 반응하는 유전자는 환경오염을 감지하는 분자마커로 개발될 수 있다. DD-PCR 기법은 차등 발현 유전자들을 발굴해내기 위한 유용한 방법으로 사용되어 왔고, 본 연구는 이 방법을 이용하여 벤조피렌에 반응하는 조피볼락 유전자들의 발굴을 목적으로진행되었다. 간조직에서 추출한 RNA로부터 벤조피렌의 노출에 의해 발현 양이 달라진 12개의 클론을 발굴하였고, 그 염기서열을 분석하였다. 또한 벤조피렌의 노출시간을 각각 6, 12, 24시간으로 달리한 조피볼락에서 12개의 클론 중 4개의 클론에 대해 northern blot 분석이 실시되었으며, 이들 모두 노출시간에 따. 본 연구결과는 오염물질의 영향에 의한 유전자들의발현에 관한 전반적인 지식을 제공하였고, 나아가 환경오염이나 외부 스트레스를 감지해 낼 수 있는 바이오마커의 개발을 위한 첫 단계로서의 정보를 제공할 수 있을 것으로 생각된다.
Nucleotide Sequence of B-tubulin Gene from the Soft Coral Scleronephthya gracillimum (Kukenthal)
염승식,우선옥,Jun-Im Song,Man Chang,Taek-kyun Lee 한국해양과학기술원 2005 Ocean science journal Vol.40 No.1
cloned the complete cDNA of the -tubulin from the soft coral, Scleronephthya gracillimum (Kkenthal) (Alcyonacea, Octocorallia, Anthozoa, Cnidaria), via the random sequencing of a cDNA library and the 5-rapid amplification of cDNA end (RACE) technique. The full-length cDNA of the S. gracillimum -tubulin comprised 1541 bp, not including the poly (A)+ stretch, also contained a complete open reading frame, which codes for a total of 445 amino acids. The amino acid residues 16-402 appeared to be in a state of conservation in a variety of animals. Northern blot analysis clearly demonstrated that the sequence we have obtained is, indeed, the full-length cDNA of the -tubulin gene in S. gracillimum.
Ubiquitin Expression in Soft Coral (Scleronephthya gracillimum) Exposed to Environmental Stresses
염승식 한국유전학회 2006 Genes & Genomics Vol.28 No.2
A cDNA clone encoding ubiquitin was isolated from the cDNA library of the soft coral Scleronephthya gracillimum, and changes in transcription of the ubiquitin gene induced by exposure to persistent organic pollutants and thermal stress were investigated using real-time quantitative polymerase reaction analysis. The S. gracillimum ubiquitin gene spanned 384 bp and encoded 128 amino acids that exhibited 96% identity and 98% positives with ubiquitin in Acropora millepora or Drosophila melanogaster and 94% identity and 96% positives with human ubiquitin. Expression of the ubiquitin gene in S. gracillimum was up-regulated approximately 4- to 7-fold during exposure to polycyclic aromatic hydrocarbons and was elevated significantly by thermal stress (28oC) compared with control (22oC) (P < 0.05). The stress-associated transcriptional changes indicated that the cellular defenses against environmental changes were activated in those animals through the up- or down-regulation of gene expression. The results also suggest that ubiquitin gene expression in response to stress may serve as a biomarker for monitoring levels of environmental contamination and the health status of animals in the local marine ecosystem.
이건섭,박소윤,염승식,우선옥,이연호,황승용,박흥식,모상현,이석찬,이택견 한국바이오칩학회 2012 BioChip Journal Vol.6 No.4
Countless species occur in the marine microalgal domain. Some are used as health functional foods or medical products but many species are harmful such as those that cause the red tide. Therefore, it is necessary to conduct prompt and accurate identification of microalgal species. As it is quite difficult to accurately distinguish all species in terms of morphology, we performed DNA barcoding analysis using molecular markers for more accurate and rapid screening. DNA barcoding analysis, i.e., DNA chip technology, is a powerful method for studies on microalgal taxonomy and biodiversity. We used the mitochondrial cytochrome c oxidase subunit I (mtCOI) as a barcoding gene to identify microalgal species. In this study, the diversity and phylogenetic differences among different microalgae were analyzed. Additionally, a microalgal species-specific probe was screened by 21-23 bp and the result was printed on silylated slide for use in a robotic microarrayer. As a result, we performed a DNA chip assay for each of 25 microalgal species and determined that the COI barcode gene was suitable as a marker gene, as it could identify various microalgae from the Korean South Sea by species.
Karyotypes of Two Ischnochiton Species : Ischnochitonidae: Polyplacophora
Choe, Byung Lae,Rho, Boon Jo,Song, Jun-Im,Yum, Seungshic 이화여자대학교 생명과학연구소 1995 생명과학연구논문집 Vol.6 No.-
연두군부(Ischnochiton comptus)와 가는줄연두군부(I. hakodadensis) 두 종의 정소를 재료로 하여 핵형 분석을 실시하였다. 두 종의 염색체는 모두 중부염색체로 구성되어 있고, 염색체 수는 n=12이며, 성염색체는 관찰되지 않았다. 이들 염색체는 이들의 크기로는 별도의 염색체군으로 나누어지지 않았다. Chromosomes prepared from the male gonads of the two polyplacophoran species. Ischnochiton comptus and I. hakodadensis were analysed. The numbers of chromosomes of the two species were 24 in diploid and 12 in haploid. In the case of I. comptus. this chromosome count coincides with the report of Nishikawa and Ishida(1969). The sex-dete-mining chromosomes have not been detected. All of the chfomosomes were classified of metacintric and could not be grouped into different classes according to their lengths.
김연정,이나영,우선옥,류재천,염승식 대한독성 유전단백체 학회 2016 Molecular & cellular toxicology Vol.12 No.4
We evaluated cadmium (Cd)-induced acute toxicity in Oryzias javanicus (marine medaka or Javanese ricefish) and gathered transcriptomic evidence for the Cd-induced endocrine-disrupting effect. The median lethal concentrations for the fish were determined to be 44.25 and 27.80 mg/L after exposure to Cd in seawater for 24 and 48 h, respectively, and 2.84, 1.61, and 1.20 mg/L after exposure in freshwater for 24, 48, and 72 h, respectively. The differences in the bioavailability and activity of free Cd2+ caused by the salt concentration in seawater could explain these dramatic differences in the toxicity of Cd between marine and fresh water system. The genes differentially expressed in O. javanicus liver tissue after exposure to 280 μg/L CdCl2 for 48 h were profiled with a customized marine medaka cDNA microarray (HazChem Fish Array). We identified 204 differentially expressed genes; the expression of 66 genes was upregulated and that of 138 genes was downregulated (P<0.05). The total 31 genes were commonly expressed in fish exposed to Cd and two references of environmental disruptor (bisphenol A, or 17β-estradiol). These genes were used to predict the changes that occur in metabolic pathways and processes in response to Cd exposure. The database for annotation, visualization and integrated discovery (DAVID) was used for functional analysis for the differentially expressed genes. Significant changes were predicted in the steroid hormone and estrogen stimulus response, vitellogenin expression, sterol and cholesterol metabolic processes, lipid transport activity, defense response, innate immune response, and metal ion binding activity. These results extend our knowledge of the toxicity of Cd at the molecular level and indicate that Cd exposure causes endocrine disruption in aquatic organisms.
김보미,조예진,이나윤,이나영,우선옥,이재성,염승식 한국바이오칩학회 2018 BioChip Journal Vol.12 No.1
While environmental concentration of bisphenol A (BPA) is ubiquitously detected in effluent and surface of freshwater, BPA has also been measured in coastal and marine ecosystems. To understand the effect of waterborne BPA on the transcriptome over time, male fish of the marine medaka Oryzias javanicus were exposed to 76 μg/L of BPA for 72 h. Time-course microarray study identified differentially expressed transcripts upon BPA exposure in the liver tissues in a time-dependent manner. The primary effect of BPA treatment appears to be significantly upregulated mRNA expression of lipid metabolism (e.g. fatty acid process, lipid/lipoprotein biosynthesis and transport, cholesterol regulation), while genes involved in diverse biochemical and physiological processes were downregulated. In particular, vertebrate immunity relevant genes showed strong downregulated patterns across time-courses. This study provides preliminary insights into the transcriptional response of marine medaka male fish to waterborne BPA exposure and suggests a basis for uncovering the mode of action particularly with respect to endocrine disrupting chemicals (EDCs) such as BPA.
우선옥,윤문근,서동상,송준임,김정하,염승식 한국유전학회 2004 Genes & Genomics Vol.26 No.4
The complete cDNA of eukaryotic translation elongation factor 1A (eEF1A) from the soft coral Scleronephthya gracillimum (Kükenthal) (Alcyonacea, Octocorallia, Anthozoa, Cnidaria) was cloned by RT-PCR and the nucleotide sequences were determined. The information on primary structure of the eEF1A gene and protein may be useful for the gene expression study as well as for the phylogenetic study. The full-length cDNA of S. gracillimum eEF1A comprised 1612 bp, not including the poly (A)+ stretch, and contained a complete open reading frame encoding 461 amino acids. The deduced amino acid sequence showed 85% identity to placozoan, 86% to human and 77% to hydra. Considering that functional domains are highly conserved throughout the evolutionary history, the present study suggests that the amino acid sequence of eEF1A alone may not resolve the phylogenetic relationship of lower metazoans. A possible solution to overcome the long branch problems that appear in cnidiarian phylogeny is discussed.