• 한국발생생물학회지:발생과생식
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• 제4권2호
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• pp.221-229
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• 2000

유선에서 젖의 생산은 뇌하수체 호르몬인 성장 호르몬과 프롤락틴을 포함한 여러 가지 호르몬의 조절을 받는다. 최근의 연구에 따르면 이 호르몬들 중에서 성장호르몬과 프롤락틴은 유선에서도 그 유전자 전사체가 발견된다 본 연구에서는 유선에서 발현되는 성장호르몬이 유선 특이 발현 유전자의 발현에 미치는 영향을 조사하고자 유선 특이 발현 유전자인 베타-락토글로불린($\beta$-lactoglobulin :BLG)의 프로모터를 모델 시스템으로 하여 소와 사람의 성장 호르몬이 유선의 유전자 발현에 끼치는 영향을 조사하였다. 성장 호르몬은 단독으로 처리하였을 패 베타-락토글로불린 유전자 프로모터 활성을 억제하였다. 그러나 젖 분비 호르몬들인 인슐린, 프롤락틴, 글루코코르티코이드와 함께 처리하였을 때는 농도 의존적으로 BLG 프로모터 활성을 상승시키는 효과를 보였다. 성장 호르몬을 유선 세포내에서 발현시켰을때는 적정농도에서 세포 증식과 유선 프로모터 활성을 크게 증진시켰다. 반면 소의 성장 호르몬 유전자 프로모터는 유선 세포에서 뚜렷한 활성을 나타내지 않았다. 이상의 결과는 유선에서 발현되는 뇌하수체 호르몬들은 조절 누수에 의한 유전자 발현이 아니라 생리적 기능을 가지고 있음을 의미한다. 또 인위적으로 성장호르몬의 발현을 조절하여 적정한 양이 발현되도록 하면 젖의 생산을 증진시킬 수 있다는 가능성도 암시한다.

• Journal of Plant Biotechnology
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• 제1권1호
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• pp.46-55
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• 1999

The expressed sequence tags(ESTs) from immature seed of rice, Oryza sativa cv Milyang 23, were partially sequenced and analyzed by homology. As of 1998, the partial sequences of about 6,600 cDNA clones were analyzed from normal and normalized immature seed cDNA libraries. About 2,200 ESTs were putatively identified by BLASTX deduced amino acid sequence homology analysis. About 20% of them were putatively identified as storage proteins. Also the clones were highly homologous to genes involved particularly in starch biosynthesis, glycolysis, signal transduction and defenses. Compared to 35% of redundancy in the ESTs of normal cDNA library, that from the substracted library was 15%. The Korea Rice Genome Network is maintained to provide the updated information of sequences, their homologies and sequence alignments of ESTs. For the stable expression of transgene in rice, diverse vectors were developed for overexpression, targeting and gene dosage effect with transit peptides (Tp) and matrix attachment region (MAR) sequence from chicken lysozyme locus. The rice calli were transformed via Agrobacterium tumefaciens LBA4404(pSB1) with the triparental mating technique and selected by herbicide resistance. The green fluorescent protein(GFP) gene in expression vector under the control of rbcS promoter-Tp was overexpressed upto 10 % of the total soluble protein. In addition, the Tp-sGFP fusion protein was properly processed during translocation into chloroplast. The expression of sGFP in the presence of MAR sequences was analyzed with Northern and immunoblot analysis. All the lines in which sGFP transgene with MAR sequence, showed position independent and copy number-dependent expression, while the lines without MAR showed the varied level of expression with the integration site. Thus the MAR sequence significantly reduced the variation in transgene expression between independent transformants.

• KSBB Journal
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• 제20권6호
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• pp.418-424
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• 2005

Rhodobacter sphaeroides는 photoheterotrophic 조건에서 성장을 하면서 질소고정효소인 nitrogenase에 의해 수소를 발생시킨다. 이 nitrogenase는 ammonium ion이 존재할 때, PII 계열 단백질인 GlnB와 GlnK에 의해 negative regulation을 받게 된다. 본 연구에서는 glnB와 glnK 유전자를 interruption하여 수소생성의 증진을 꾀하였다. 그 결과, parental strain이나 glnB 혹은 glnK 유전자 하나만을 interruption 시켰을 경우, 암모니움 이온에 의해 nitrogenase 활성이 저해를 받고 있었으나, glnB, glnK 모두를 interruption 시켰을 때, 암모니움 이온에 의한 저해 효과가 줄어드는 것을 볼 수 있었다. 또한 이러한 glnB-glnK double mutant에 nifDK gene을 in trans로 넣어 nitrogenase의 dosage를 높였을 경우, 수소 생성이 약 30% 가량 증진되었다는 것을 볼 수 있었다.

• Journal of Genetic Medicine
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• 제13권2호
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• pp.78-88
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• 2016

Purpose: To identify the clinical characteristics of SRY-negative male patients and genes related to male sex reversal, we performed a retrospective study using cases of 46,XX testicular disorders of sex development with a review of the literature. Materials and Methods:SRY-negative cases of 46,XX testicular disorders of sex development referred for cytogenetic analysis from 1983 to 2013 were examined using clinical findings, seminal analyses, basal hormone profiles, conventional cytogenetic analysis and polymerase chain reaction. Results: Chromosome analysis of cultured peripheral blood cells of 8,386 individuals found 19 cases (0.23%) with 46,XX testicular disorders of sex development. The SRY gene was confirmed to be absent in three of these 19 cases (15.8%). Conclusion: We report three rare cases of SRY-negative 46,XX testicular disorders of sex development. Genes on autosomes and the X chromosome that may have a role in sex determination were deduced through a literature review. These genes, through differences in gene dosage variation, may have a role in sex reversal in the absence of SRY.

• The Korean Journal of Physiology and Pharmacology
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• 제27권1호
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• pp.31-38
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• 2023

Carboplatin, an advanced anticancer drug with excellent efficacy against ovarian cancer, was developed to alleviate the side effects that often occur with cisplatin and other platinum-based compounds. Our study reports the in vitro characteristics, viability, and activity of cells expressing the inducible nitric oxide synthase (iNOS) gene after carboplatin was conjugated with polysuccinimide (PSI) and administered in combination with other widely used anticancer drugs. PSI, which has promising properties as a drug delivery material, could provide a platform for prolonging carboplatin release, regulating its dosage, and improving its side effects. The iNOS gene has been shown to play an important role in both cancer cell survival and inhibition. Herein, we synthesized a PSI-carboplatin conjugate to create a modified anticancer agent and confirmed its successful conjugation. To ensure its solubility in water, we further modified the structure of the PSI-carboplatin conjugate with 2-aminoethanol groups. To validate its biological characteristics, the ovarian cancer cell line SKOV-3 and normal ovarian Chinese hamster ovary cells were treated with the PSI-carboplatin conjugate alone and in combination with paclitaxel and topotecan, both of which are used in conventional chemotherapy. Notably, PSI-carboplatin conjugation can be used to predict changes in the genes involved in cancer growth and inhibition. In conclusion, combination treatment with the newly synthesized polymer-carboplatin conjugate and paclitaxel displayed anticancer activity against ovarian cancer cells but was not toxic to normal ovarian cancer cells, resulting in the development of an effective candidate anticancer drug without severe side effects.

• Animal Bioscience
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• 제36권8호
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• pp.1285-1292
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• 2023

Objective: The objective of this study was to develop a novel endolysin (PanLys.1) for the specific killing of the ruminal hyper-ammonia-producing bacterium Peptostreptococcus anaerobius (P. anaerobius). Methods: Whole genome sequences of P. anaerobius strains and related bacteriophages were collected from the National Center for Biotechnology Information database, and the candidate gene for PanLys.1 was isolated based on amino acid sequences and conserved domain database (CDD) analysis. The gene was overexpressed using a pET system in Escherichia coli BL21 (DE3). The lytic activity of PanLys.1 was evaluated under various conditions (dosage, pH, temperature, NaCl, and metal ions) to determine the optimal lytic activity conditions. Finally, the killing activity of PanLys.1 against P. anaerobius was confirmed using an in vitro rumen fermentation system. Results: CDD analysis showed that PanLys.1 has a modular design with a catalytic domain, amidase-2, at the N-terminal, and a cell wall binding domain, from the CW-7 superfamily, at the C-terminal. The lytic activity of PanLys.1 against P. anaerobius was the highest at pH 8.0 (p<0.05) and was maintained at 37℃ to 45℃, and 0 to 250 mM NaCl. The activity of PanLys.1 significantly decreased (p<0.05) after Mn²⁺ or Zn²⁺ treatment. The relative abundance of P. anaerobius did not decrease after administration PanLys.1 under in vitro rumen conditions. Conclusion: The application of PanLys.1 to modulate P. anaerobius in the rumen might not be feasible because its lytic activity was not observed in in vitro rumen system.

• Clinical and Experimental Reproductive Medicine
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• 제37권1호
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• pp.25-31
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• 2010

목 적: X 염색체 불활성화는 여성과 남성 사이에 X 염색체의 유전자 발현 유지를 위해 여성의 X 염색체 중 하나가 불활성화 되는 현상이다. 이러한 X 염색체 불활성화는 해독되지 않는 XIST 유전자에 의해 조절된다. XIST 유전자는 오직 불활성화된 X 염색체 에서만 발현되고, 활성화된 X 염색체 에서는 발현되지 않는다. 따라서 체세포에서 활성화된 X 염색체의 XIST 유전자는 promoter 부분이 메틸화 되어있고, 불활성화된 X 염색체에서는 메틸화가 거의 되어 있지 않다. 연구방법: 본 연구에서는 정상 여성과 정상 남성의 XIST 유전자의 promoter와 5'-end 지역의 메틸화 차이를 측정하기 위해 정상여성과 남성의 혈액에서 DNA를 추출하여 파이로시퀀싱 (Pyrosequencing) 방법을 통해 XIST 유전자의 총 8부분의 CpG 영역 (-1696, -1679, -1475, -1473, -1469, +947, +956, +971)을 분석하였다. 결 과: 총 8부분의 CpG 영역을 분석한 결과, promoter 부분인 CpG 1-5 영역 (-1696, -1679, -1475, -1473, -1469)에서는 여성과 남성의 메틸화 정도에 차이가 없었다. 그러나 5'-end 부분인 CpG6-8 영역 (+947, +956, +971)에서는 여성이 45.2% 49.9% 44.2%, 남성이 90.6%, 96.7%, 87.8%으로 메틸화 정도가 차이를 나타냈다. 결 론: 따라서 본 연구에 사용한 방법은 XIST 유전자의 메틸화 패턴의 차이를 기존의 방법보다 신속하고 정확하게 분석할 수 있다는 장점이 있기 때문에 유용하게 사용될 수 있을 것이다.

• BMB Reports
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• 제32권1호
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• pp.92-97
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• 1999

Cytokinesis and septation should be coordinated to nuclear division in the cell division cycle for precise transmission of the genome into daughter cells. byr4, an essential gene in fission yeast Schizosaccharomyces pombe, regulates the timing of cytokinesis and septation in a dosage-dependent manner. We examined the intracellular localization of the Byr4 protein by expressing byr4 as a fusion of green fluorescence protein (GFP). The Byr4 protein localizes as a single dot on the nuclear periphery of interphase cells, duplicates before mitosis, and the duplicated dots segregate with the nuclei in anaphase. The behavior of Byr4p throughout the cell cycle strongly suggests that Byr4p is localized to the spindle pole body (SPB), a microtubule organizing center (MTOC) in yeast. The presence of the Byr4 protein in the SPB is consistent with its function to coordinate mitosis and cytokinesis. We also mapped the domains of Byr4p for its proper localization to SPB by expressing various byr4 deletion mutants as GFP fusions. Analyses of the diverse byr4 deletion mutants suggest that the indirect repeats and the regions homologous to the open reading frame (ORF) YJR053W of S. cerevisiae in its C-terminus are essential for its localization to the SPB.

• KSBB Journal
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• 제29권4호
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• pp.263-270
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• 2014

This study reports significant improvement of atopic dermatitis condition as a result of experiment using Xanthium strumarium L. extract (XS-E) at the dorsal skin of induced atopic dermatitis Nc/Nga mice. Skin clinical score has decreased ($2.75{\pm}0.85$, *p<0.05), showing visible change of skin condition. IgE (***p<0.001) and IgG1 ($2522.00{\pm}32.80$, ***p<0.001) in plasma also decreased significantly. mRNA (gene expression) level increased ($RQ=2.75{\pm}0.10$, ***p<0.001) within skin tissue of CD4+CD25+Foxp3+ Treg cell that's activated by XS-E dosage, thereby discovering that there is an effect of suppressing proliferation and viability of Th2 cell, eosinophils, mast cell and inflammatory cell. Upon examining cells permeated with H&E and toluidine blue staining technique, thickness of epidermis and mast cell's permeation decreased, and the result of examining the distribution of CCR 3+ eosinophils within ALN showed that it's level fell down to that of wild type (normal group, NC/Nga-WT). By such results, it is suggested that XS-E is highly effective on atopic dermatitis, and it is considered that continued quantitative research and case study of clinical research such as effect of cell number in individual tissues or change of total cell number are necessary.

• Molecules and Cells
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• 제23권3호
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• pp.331-339
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• 2007

It has been suggested that the structure and function of nuclear receptors are evolutionally conserved. Here, we compare the molecular functions of the nile tilapia (Oreochromis niloticus) small heterodimer partner (nSHP/NR0B2) and the Dosage-sensitive sex reversal AHC critical region on X chromosome gene 1 (nDAX-1/NR0B1) with those of human SHP and DAX-1 (hSHP and hDAX-1, respectively). We found that, upon transient cotransfection of human cells, nDAX-1 repressed the activity of tilapia SF-1 (nSF-1) but not that of human SF-1, although the physical interaction with human SF-1 was retained. Similarly, nSHP repressed the activity of nSF-1, whereas hSHP did not, pointing to divergent evolution of SHP/SF-1 in fish and human. We thus propose that the repressive functions of SHP and DAX-1 have been conserved in fish and mammals although with different transcriptional targets and mechanisms. These differences provide new insights into the physiological diversification of atypical orphan nuclear receptors during vertebrate evolution.