• Title/Summary/Keyword: MiRNA

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Isolation and Characterization of a Chitin Synthase Gene Fragments from Pleurotus sajor-caju (여름느타리의 Chitin synthase 유전자 단편분리 및 발현 특성 분석)

  • Jeong, Mi-Jeong;Park, Soo-Chul;Kim, Bum-Gi;Yoo, Young-Bok;Ryu, Jin-Chang
    • The Korean Journal of Mycology
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    • v.26 no.3 s.86
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    • pp.354-360
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    • 1998
  • We isolated three amplified DNA fragments from P. sajor-caju by Polmerase chain reaction (PCR) using the chithin synthase specific primers. Since the sequence analysis of the these fragments showed significant homology to the other known chitin synthase gene, we regarded these cloned fragments as PsCHS1, PsCHS2, and PsCHS3 according to their size. The PsCHS3, which showed the highest sequence homology (83% identity in amino acid level with ChsI of Rhizopus oligosporus in conserved region), was selected to see expression pattern of the corresponding gene. The result of RT-PCR using internal primer of the PsCHS3 fragment revealed that PsCHS3 gene was only expressed in cap and mycelium but not in stipe. In order to see whether the PsCHS3 gene was to be induced by wounding, the comparison of the mRNA level of this gene between wounded and unwounded mature cap showed at least two times induction of this gene by wounding treatment.

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Investigation of the Gene Encoding Isotocin and its Expression in Cinnamon Clownfish, Amphiprion melanopus (Cinnamon clownfish Amphiprion melnaopus의 이소토신 유전자 구조와 삼투압 조절이 미치는 영향)

  • Noh, Gyeong Eon;Choi, Mi-Jin;Min, Byung Hwa;Rho, Sum;Kim, Jong-Myoung
    • Journal of Life Science
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    • v.26 no.2
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    • pp.164-173
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    • 2016
  • Isotocin (IT), a nonapeptide homolog of oxytocin in mammals, has been suggested to be involved in physiological processes including social behaviors, stress responses, and osmoregulation in teleost fish. To study its structure and function, the gene encoding the IT precursor was cloned from the genomic DNA and brain cDNA of the cinnamon clownfish, Amphiprion melanopus. The IT precursor gene consists of three exons separated by two introns, and encodes an open reading frame of 156 amino acid (aa) residues, comprising a putative signal peptide of 19 aa, a mature IT protein of 9 aa, a proteolytic processing site of 3 aa, and 125 aa of neurophysin. Tissue-specific analysis of the IT precursor transcript indicated its expression in the brain and gonads of A. melanopus. To examine its osmoregulatory effects, the salinity of the seawater (34 ppt) used for rearing A. melanopus was lowered to 15 ppt. Histological analysis of the gills indicated the apparent disappearance of an apical crypt on the surface of the gill lamella of A. melanopus, as pavement cells covered the surface upon acclimation to the lower salinity. The level of Na+/K+-ATPase activity in the gills was increased during the initial stage of acclimation, followed by a decrease to its normal level, suggesting its involvement in osmoregulation and homeostasis. The only slight increase in the level of IT precursor transcript in the A. melanopus brain upon low-salinity acclimation suggested that IT played a minor role, if any, in the process of osmoregulation.

A Study on the Sensory Characteristic of Yogurt and Antimicrobial Activity of Lactobacillus plantarum LHC52 Isolated from Kimchi (김치에서 분리한 Lactobacillus plantarum LHC52의 항균활성과 요구르트의 관능성 연구)

  • Lee, Seung-Gyu;Han, Ki-Sung;Jeong, Seok-Geun;Oh, Mi-Hwa;Jang, Ae-Ra;Kim, Dong-Hun;Bae, In-Hyu;Ham, Jun-Sang
    • Food Science of Animal Resources
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    • v.30 no.2
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    • pp.328-335
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    • 2010
  • The aim of our study was to develop a new starter culture for fermented milk. Polymerase chain reaction screening of 103 acid-producing isolates from Kimchi identified 72 Lactobacillus strains. The ability of the strains to inhibit the growth of the food-borne human pathogens (Escherichia coli, Salmonella Enteritidis, Staphylococcus aureus) was measured, using a conventional paper disk method. Among the 72 strains, strain LHC52 displayed potent antagonistic activity. Use of 16S rDNA sequencing and the API 50CHL system identified the strain as Lactobacillus plantarum and it was designated L. plantarum LHC52. Biochemical analyses revealed especially high antibacterial activity against E. coli. Yogurt produced using L. plantarum LHC52 did not show different microbiological and physicochemical properties compared to conventionally-prepared yogurt, implicating L. plantarum LHC52 as a useful, potently antibacterial starter culture for yogurt preparation.

Inhibitory Effect of Extracts from Rhododendron Brachycarpum and Abies Koreana E.H. Wilson on Degranulation and Cytokine Expression in RBL-2H3 Cells (만병초와 구상나무 추출물의 RBL-2H3 세포 탈과립, 싸이토카인 유전자 발현에 미치는 영향)

  • Jung, Eui-Man;Kim, Jae-Woo;Park, Mi-Jin;Lee, Sung-Suk;Choi, Don-Ha;Lee, Jungbok;Jeung, Eui-Bae
    • Journal of the Korean Wood Science and Technology
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    • v.41 no.6
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    • pp.551-558
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    • 2013
  • Ethanol extracts from Rhododendrom brachycarpum and Abies koreana were investigated for their anti-allergic activities using RBL-2H3 cell line. After treatment with ethanol extracts of various concentrations on the immune response induced mast cell by concanavalin A (Con A), the expressions of cytokine interleukin-4 (IL-4), interleukin-13 (IL-13) were determined by using RT-PCR and the degranulation of mast cells was determined by measuring ${\beta}$-hexosaminidase release. Expression level of IL-4 was decreased by the extract from Rhododendrom brachycarpum in $10^{-7}$, $10^{-5}$ and $10^{-3}%$ concentrations. Expression level of IL-13 was also decreased by both extracts. ${\beta}$-Hexosaminidase release by RBL-2H3 cells was inhibited at the $10^{-5}$ and $10^{-3}%$ concentration of extracts from Rhododendrom brachycarpum and Abies koreana, respectively. These results demonstrate that ethanol extracts of Rhododendron brachycarpum and Abies koreana exert anti-allergic effects by regulating the reduction of IL-4 and IL-13 genes expression and also the secretion of ${\beta}$-hexosaminidase.

Studies on the Sugar Analysis and Biological Activity of Sap from Juglans mandshurica Maxim. (가래나무 수액의 당성분 분석 및 생리활성 연구)

  • Kim, Dae-Wook;Choi, Mi-Na;Lee, Min-Sung;Jung, Hae-Suk;Byeon, Jun-Gi;Kim, Yeong-Su
    • Journal of Chitin and Chitosan
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    • v.23 no.4
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    • pp.262-266
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    • 2018
  • Even though the saps have been consumed as beverage in Korea, however a little research has been conducted on the chemical composition. We determined free sugars, mineral in the sap from Juglans mandshurica Maxim. collected from BongHwa region. The contents of glucose, sucrose, and fructose in the sap from J. mandshurica were 0.15, 2.73, and 0.09%, respectively. The prominent minerals in the sap from J. mandshurica were calcium and potassium. The contents of Ca, K, Mg, Fe, and Mn of sap from J. mandshurica were 0.61, 0.57, 0.12, 0.002, and 0.014 ppm, respectively. The sap from J. mandshurica show inhibitory effect on elastase. And real-time RT-PCR showed that sap from J. mandshurica increased mRNA level of AQP3 and HAS2 gene and increased hyaluronic acid production in HaCaT cells. These results indicated that the sap from J. mandshurica can potentially be used for developing cosmetic ingredient for skin moisturizing and anti-aging.

Establishment of Human Embryonic Stem Cells using Mouse Embryonic Fibroblasts and Human Fetal Fibroblasts as Feeder Cells (인간태아 섬유아세포와 생쥐배아 섬유아세포를 기저세포로 활용한 인간 배아줄기세포의 확립)

  • Cho, Hye Won;Ko, Kyoung Rae;Kim, Mi Kyoung;Lee, Jae Ik;Sin, Su Il;Lee, Dong Hyung;Kim, Ki Hyung;Lee, Kyu Sup
    • Clinical and Experimental Reproductive Medicine
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    • v.32 no.2
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    • pp.133-147
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    • 2005
  • Objectives: This study was carried out to establish human embryonic stem cells derived from frozen-thawed embryos using mouse embryonic fibroblasts (mEFs), human fetal skin and muscle fibroblasts as feeder cells, and to identify the characteristic of embryonic stem cells. Methods: When primary mEFs, human fetal skin and muscle fibroblasts were prepared, passaging on 4 days from replating could have effective trypsinization and clear feeder layers. Eight of 23 frozenthawed 4~8 cell stage embryos donated from consenting couples developed to blastocysts. Inner cell mass (ICM) was isolated by immunosurgery. ICM was co-cultured on mEFs, human fetal skin or muscle fibroblasts. The ICM colonies grown on mEFs, human fetal skin or muscle fibroblasts were tested the expression of stage specific embryonic antigen-3, -4 (SSEA-3, -4), octamer binding transcription factor-4 mRNA (Oct-4) and alkaline phosphatase surface marker. Results: We obtained 1 ICM colony from 2 ICM co-cultured on mEFs as feeder cells and did not obtain any ICM colony from 6 ICM clumps co-cultured on human fetal skin or muscle fibroblasts. The colony formed on mEFs could be passaged 30 times every 5 days with sustaining undifferentiated colony appearance. When the colonies cultured on mEFs were grown on human fetal skin or muscle fibroblasts, the colonies could be passaged 15 times every 9 days with sustaining undifferentiated colony appearance. The colonies grown on mEFs and human fetal fibroblasts expressed SSEA-4 and alkaline phosphatase surface markers and positive for the expression of Oct-4 by reverse transcription-polymerase chain reaction (RT-PCR). The produced embryoid body differentiated spontaneously to neural progenitorlike cells, neuron-like cells and beating cardiomyocyte-like cells, and frozen-thawed embryonic stem cells displayed normal 46,XX karyotype. Conclusions: The human embryonic stem cells can be established by using mEFs and human fetal fibroblasts produced in laboratory as feeder cells.

Anti-atopic Effect of Zizyphus jujuba var. spinosa Composite oil(ZjJ-C_oil) and Aralia cordata var. continentalis Composite oil(ARC-C_oil) in DNCB-induced Atopic Dermatitis-like Skin Lesions NC/Nga Mice (천연물 유래 산조인 복합오일 (Zizyphus jujuba var. spinosa Composite oil, ZjJ-C_oil) 및 독활 복합오일 (Aralia cordata var. continentalis Composite oil, ARC-C_oil)의 DNCB로 유도된 NC/Nga 생쥐의 아토피에 미치는 효과 연구)

  • Kwak, Jin-young;Choir, Hee-Jeong;Park, Jung-mi;Park, Jung-hwan;Kho, Young-mee;Jang, Tae-soo;Ahn, Teakwon
    • Journal of Sasang Constitutional Medicine
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    • v.29 no.4
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    • pp.347-368
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    • 2017
  • The aim of this study is to investigate the effect of Zizyphus jujuba var. spinosa Composite oil(ZjJ-C_oil) and Aralia cordata var. continentalis Composite oil(ARC-C_oil) to NC/Nga mice induced in Atopic dermatitis-like skin lesions by DNCB. NC/Nga mice which have been induced to Atopic dermatitis-like skin lesions by DNCB are divided into 4 groups, the first is the mice which have been spread with izyphus jujuba var. spinosa Composite oil(ZjJ-C_oil), the second is the mice which have been spread with Aralia cordata var. continentalis Composite oil(ARC-C_oil), the third is which have been spread with dexamethasone (Dexa.) 0.5% on their Atopic lesion, the last is the control group. Then We analyzed skin clinical score, blood sample of each group of measure state of the dorsal skin, the number of immunocytes, and resect the skin lesion to anlayze the state of cells. There are meaningful results of measuring the number of IgE, IL-4, IL-13, $IFN-{\gamma}$, IL-5, the total cells in ALN, dorsal skin, CD4+ Th, CD11b+/Gr-1+ in PBMCs, CD4+ Th, B220+/CD23+ in ALN, $CD4^+$, $CD8^+$ Th in dorsal skin, the level of COX-2, $TNF-{\alpha}$, $TGF-{\beta}1$, IL-4, IL-13 mRNA, the state of the skin lesion and cells in the group with ZjJ-C_oil, ARC-C_oil cream in comprarison with the control group.

Inhibitory Effects of Tannic acid on Human Skin Fibroblast Elastase Activity (사람의 섬유아세포 엘라스타제 활성에 대한 탄닌산의 억제 효과)

  • Lee, Ju-Eun;Kim, So-Young;Kim, Su-Yeon;Oh, Mi-Hee;Yun, Hye-Young;Baek, Kwang-Jin;Kwon, Nyoun-Soo;Kim, Dong-Seok
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.34 no.3
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    • pp.217-223
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    • 2008
  • Elastin is an important component of elastic fibers in the skin. Recently, many studies have reported that elastin is also involved In inhibiting or repairing wrinkle formation, although collagen is a major factor in the skin wrinkle formation. Elastase is a metalloproteinase which acts on degradation of elastin. It is known that elastase activity is increased by ultraviolet (UV) B radiation. Thus, Increased elastase activity could be the major reason for skin elasticity reduction and winkle formation. Tannic acid is a polyphenol found in various fruits and nuts. This molecule has a potent ability to eliminate reactive oxygen species and reactive nitrogen species. In the present study, we investigated whether tannic acid has effects on elastase activity and tropoelastin synthesis. Our results showed that tannic acid reduced elastase activity significantly in a dose-dependent manner. However, the expression of tropoelastin protein and mRNA was not significantly affected by tannic acid. From these results, we suggest that tannic acid may contribute to block tortuosity of elastic fibers by inhibiting elastase. Thus, tannic acid might be developed for a possible agent to Inhibit skin aging.

In Vitro Tissue Culture Frequency and Transformation of Various Cultivars of Soybean (Glycine max (L.) Merr.) (다양한 콩 자원들의 기내 조직배양 효율 및 형질전환)

  • Seo, Mi-Suk;Cho, Chuloh;Jeong, Namhee;Sung, Soon-Kee;Choi, Man-Soo;Jin, Mina;Kim, Dool-Yi
    • Korean Journal of Plant Resources
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    • v.34 no.4
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    • pp.278-286
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    • 2021
  • Efficient in vitro regeneration system is essential for the successful crop breeding of soybean (Glycine max (L.) Merr.) using the new biotechnology. The genotype of donor plants strongly influences the establishment of tissue culture system. Therefore, the screening of genotypes with excellent tissue culture ability is very important for soybean genetic improvement. In this study, we report the tissue culture efficiency of 21 soybean cultivars belong to Korean soybean core-collection and two foreign cultivars (Jack and Maverick). The Kwangan, Anpyeong and Seonam are share close genetic relationship in 21 cultivars and these three cultivars were observed the high frequency of germination and regeneration. Furthermore, the high tissue culture abilities were also observed in the Williams 82 used in reference genome sequencing and the two foreign cultivars. The transformation of pBAtc:tRNA with bar gene was performed by Agrobacterium tumefaciens in the cultivars with high tissue culture ability. Transformation of the bar gene was identified by PCR analysis in Kwangan, Pungwon, Seonam, and Maverick. Our results provide useful information for the breeding of various soybean cultivars by plant biotechnology such as, genome editing.

Inhibitory Effect of the Branch Extracts from Taxillus yadoriki Parasitic to Neolitsea sericea against the Cell Proliferation in Human Lung Cancer Cells, A549 (참식나무(Neolitsea sericea) 기주 참나무겨우살이(Taxillus yadoriki) 가지 추출물의 폐암세포 A549에 대한 세포생육 억제활성)

  • Park, Su Bin;Kim, Ha Na;Kim, Jeong Dong;Park, Gwang Hun;Eo, Hyun Ji;An, Mi-Yun;Jeong, Jin Boo
    • Korean Journal of Plant Resources
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    • v.32 no.2
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    • pp.109-115
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    • 2019
  • In this study, we evaluated the anti-cancer activity and potential molecular mechanism of 70% ethanol extracts of branches from Taxillus yadoriki parasitic to Neolitsea sericea (TN-NS-B) against human lung cancer cells, A549. TY-NS-B dose-dependently suppressed the growth of A549 cells. TY-NS-B decreased ${\beta}$-catenin protein level, but not mRNA level in A549 cells. The downregulation of ${\beta}$-catenin protein level by TY-NS-B was attenuated in the presence of MG132. Although TY-NS-B phosphorylated ${\beta}$-catenin protein, the inhibition of $GSK3{\beta}$ by LiCl did not blocked the reduction of ${\beta}$-catenin by TY-NS-B. In addition, TY-NS-B decreased ${\beta}$-catenin protein in A549 cells transfected with Flag-tagged wild type ${\beta}$-catenin or Flag-tagged S33/S37/T41 mutant ${\beta}$-catenin construct. Our results suggested that TN-NS-B may downregulate ${\beta}$-catenin protein level independent on $GSK3{\beta}$-induced ${\beta}$-catenin phosphorylation. Based on these findings, TY-NS-B may be a potential candidate for the development of chemopreventive or therapeutic agents for human lung cancer.