• Title/Summary/Keyword: Gene analysis

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Microbial Diversity in the Enrichment Cultures from the Fermented Beverage of Plant Extract Using Ribosomal RNA Sequence Analysis (라이보좀 RNA 염기서열 분석을 이용한 집식배양된 식물추출물발효음료의 미생물 다양성)

  • Lee, Choung Kyu;Kim, Baolo;Kang, Young Min;Lee, Hee Yul;Hwang, Chung Eun;Ahn, Min Ju;Seo, Weon Taek;Cho, Kye Man
    • Korean Journal of Microbiology
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    • v.50 no.4
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    • pp.351-359
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    • 2014
  • A beverage was produced by the fermentation of mixed extracts from the various fruits, vegetables, algae, and medical herbs. The physicochemical properties of the fermented beverage of plant extracts (FBPE) and microbial diversity were analyzed in cultures enriched from FBPE using 16S and 26S rRNA gene sequence analyses. The pH, acidity, $^{\circ}brix$, reducing sugar, and alcohol contents of the FBPE were determined to be the 3.48, 1.68%, 70.0, 1,026 g/L, and 3.5%, respectively. The most abundant free sugar and organic acid in the FBPE were glucose (567.83 g/L) and tartaric acid (93.68 mg/L), respectively. Lactobacillus homohiochii was the predominant species in all enriched culture samples: 100% of the species in 0B (0% sugar) and 40B (40% sugar) libraries and 95.6% of 20B library (20% sugar). Lactobacillus fructivorans was detected in the 20B library. The predominant species in the samples of enrichment cultures collected from FBPE with three different sugar concentrations were: Candida zeylanoides (45.2%) in the 0Y library (0% sugar), Candida lactis-condensi (35.7%) and C. zeylanoides (35.7%) in the 20Y library (20% sugar), and C. lactis-condensi (38.1%) in the 40Y library (40% sugar). This result may provide a useful frame of reference for further analyses of microbial population dynamics in FBPE.

Anti-atopic Effects of Castanea crenata Inner Shell Extracts Fermented by Lactobacillus bifermentans (Lactobacillus bifermentans로 발효한 율피의 항아토피 효과)

  • Kim, Bae Jin;Son, Woo Rim;Choi, Mi Ok;Jo, Seung Kyeung;Jung, Hee Kyoung;Lee, Jin Tae;Kim, Hak Yoon;Kwoen, Dae Jun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.9
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    • pp.1378-1386
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    • 2013
  • Atopic dermatitis (AD) is a common skin disease characterized by chronic and relapsing inflammatory dermatitis with immunological disturbances. In spite of the continuous increase in the incidence of AD, it is regrettable that till date there is no effective treatment to treat the same. Therefore, the present study was designed to examine the possible anti-atopic effects of Castanea crenata inner shell extracts fermented by Lactobacillus bifermentans (FCS) in 2,4-dinitrochlorobenzene (DNCB) induced AD in NC/Nga mice. Based on the results of HPLC analysis, we found that FCS contains anti-inflammatory factors such as gallic acid (10.18 mg/g) and ellagic acid (2.14 mg/g). The groups that we have used in this study included 0.1%, 1%, 5% fermented Castanea crenata inner shell extracts (FCS 0.1, FCS 1, FCS 5), 1,3-butylene glycol treated control (AD), and normal mice. After topical FCS treatment, we observed that the clinical severity score for AD was lower in both the FCS 1 and FCS 5 groups than the AD group. We also proved beyond doubt that there was improvement of melanin, erythema and skin moisture indices in the FCS 5 group. Spleen index and gene expression levels of pro-inflammatory cytokines such as IL-$1{\beta}$ and TNF-${\alpha}$ were significantly decreased in the FCS 5 group compared to the AD group (P<0.05). Further, we also found that the level of serum immunoglobulin E (IgE) in the FCS-treated group was decreased in a concentration-dependent manner. The results of our study suggest that FCS can be effectively used as a cosmeceutical ingredient for both the prevention and improvement of AD.

Molecular Cytogenetic Characterization of Supernumerary Marker Chromosomes by Chromosomal Microarray (염색체 마이크로어레이를 이용한 표지염색체의 분자세포유전학적 특성)

  • Bae, Mi-Hyun;Yoo, Han-Wook;Lee, Jin-Ok;Hong, Maria;Seo, Eul-Ju
    • Journal of Genetic Medicine
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    • v.8 no.2
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    • pp.119-124
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    • 2011
  • Purpose: Supernumerary marker chromosome (SMC) could be associated with various phenotypic abnormalities based on the chromosomal origin of SMCs. The present study aimed to determine the genomic contents of SMCs using chromosomal microarray and to analyze molecular cytogenetic characterizations and clinical phenotypes in patients with SMCs. Materials and Methods: Among patients with SMCs detected in routine chromosomal analysis, SMCs originating from chromosome 15 were excluded from the present study. CGH-based oligonucleotide chromosomal microarray was performed in 4 patients. Results: The chromosomal origins of SMCs were identified in 3 patients. Case 1 had a SMC of 16.1 Mb in 1q21.1-q23.3. Case 2 showed 21 Mb gain in 19p13.11-q13.12. Case 3 had a 4.5 Mb-sized SMC rearranged from 2 regions of 2.5 Mb in 22q11.1-q11.21 and 2.0 Mb in 22q11.22-q11.23. Conclusion: Case 1 presented a wide range of phenotypic abnormalities including the phenotype of 1q21.1 duplication syndrome. In case 2, Asperger-like symptoms are apparently related to 19p12-q13.11, hearing problems and strabismus to 19p13.11 and other features to 19q13.12. Compared with cat-eye syndrome type I and 22q11.2 microduplication syndrome, anal atresia in case 3 is likely related to 22q11.1-q11.21 while other features are related to 22q11.22-q11.23. Analyzing SMCs using high-resolution chromosomal microarray can help identify specific gene contents and to offer proper genetic counseling by determining genotype-phenotype correlations.

Genetic Polymorphism in Corticotropin-releasing Hormone Receptor Type-1 in Preeclamptic Korean Women

  • Lim, Ji-Hyae;Kim, Shin-Young;Park, So-Yeon;Kim, Do-Jin;Kim, Mi-Jin;Ahn, Hyun-Kyong;Han, Jung-Yeol;Kim, Moon-Young;Park, Hyun-Young;Lee, Kwang-Soo;Kim, Young-Ju;Ryu, Hyun-Mee
    • Journal of Genetic Medicine
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    • v.8 no.2
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    • pp.113-118
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    • 2011
  • Purpose: Placental corticotropin-releasing hormone receptor type 1 (CRHR1) expression is reduced in pregnancies with abnormal placental function such as preeclampsia (PE), and the levels and/or function of CRHR1 are genetically influenced. The aim of this study was to investigate the association between the c.33+8199C>T polymorphism in the CRHR1 gene and PE in a Korean population. Materials and Methods: Using a case-control design, the association between the CRHR1 polymorphism and the risk of PE was investigated in 203 individuals with PE and 211 normotensive controls. Genotypes were determined using a SNapShot kit and an ABI Prism 3100 Genetic analyzer. Results: Genotypes and allele frequencies for the CRHR1 polymorphism did not differ between PE and normotensive pregnancies. The variant T allele was more frequent than the ancestral C allele in both of the groups and was more frequent in the controls than in the cases. In risk analysis for PE, there was not an increased risk of preeclampsia in subjects who were concomitant homozygous rare allele genotypes (CC) (OR, 0.3; P=0.15) or heterozygous rare allele genotypes (TC) (OR, 0.8; P=0.29). There were no differences in the complications of PE such as severity or preterm delivery in patients with the CRHR1 polymorphism. Conclusion: Our findings indicate that the CRHR1 polymorphism was not associated with PE in the present Korean study group.

Inheritance of Kunitz Trypsin Inhibitor and P34 Protein in Soybean Seed (콩 종자에서 쿠니츠트립신인히비터와 P34 단백질의 유전)

  • Han, Eun-Hui;Sung, Mi-Kyung;Baek, Woon-Jang;Shim, Sang-In;Kim, Min-Chul;Chung, Jong-Il
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.57 no.1
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    • pp.78-82
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    • 2012
  • Soybean [$Glycine$ $max$ (L.) Merr.] protein is a high quality source for food and feed. But, antinutritional factors in the raw mature soybean are exist. Kunitz trypsin inhibitor (KTI) protein is a main antinutritional factor in soybean seed. Also, P34 protein, referred as $Gly$ $m$ Bd 30K, has been identified as a predominant immunodominant allergen. Genetic relationship between KTI protein and P34 protein could be useful in soybean breeding program for the genetic elimination or reduction of these factors. The objective of this study was to determine the independent inheritance or linkage between KTI protein and P34 protein in soybean seed. A total of 479 $F_2$ seeds were obtained from the cross of 07B1 and PI567476 parents. KTI protein and relative amount of P34 protein were analysed from $F_2$ seeds harvested from the F1 plants by using SDS-PAGE and Western blot analysis. The segregation ratios of 3 : 1 for KTI protein (353 KTI protein present : 126 KTI protein absent) and relative amount of P34 protein (363 normal amount of P34 protein : 116 low amount of P34 protein). The segregation ratio of 3 : 1 suggested that KTI protein and relative amount of P34 protein in mature soybean seed were controlled by a single major gene. The segregation ratios of 9 : 3 : 3 : 1 (266 KTI protein present, normal amount of P34 protein: 88 KTI protein present, low amount of P34 protein: 102 KTI protein absent, normal amount of P34 protein: 23 KTI protein absent, low amount of P34 protein) and Chi-square value (${\chi}^2$=3.31, P=0.346) were observed in $F_2$ seeds. This data showed that KTI protein was inherited independently with relative amount of P34 protein in soybean. These results will be helpful in breeding program for selecting the line with lacking KTI protein and reduced amount of P34 protein in soybean.

Physiological and Proteome Responses of Korean F1 maize (Zea mays L.) Hybrids to Water-deficit Stress during Tassel Initiation (옥수수 영양생장기 한발 스트레스에 의한 광합성의 생리적 반응 및 프로테옴 변화 분석)

  • Bae, Hwan Hee;Kwon, Young-Sang;Son, Beom-Young;Kim, Jung-Tae;Go, Young Sam;Kim, Sun-Lim;Baek, Seong-Bum;Shin, Seonghyu;Kim, Sang Gon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.64 no.4
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    • pp.422-431
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    • 2019
  • Severe droughts in spring have occurred frequently in Korea in recent years, exerting a critical impact on corn yield. Therefore, it is necessary to find physiological and/or molecular indicators of the response to drought stress in maize plants. In this study, we investigated the effects of water-deficit stress on two Korean elite F1 maize hybrids, Ilmichal and Gwangpyeongok, by withholding water for 10 days at tassel initiation. The water deficit drastically reduced the relative leaf water content, leaf number, leaf area, and stem length, leading to dry matter reduction. Moreover, it reduced the SPAD values and stomatal conductance of leaves in drought-stressed plants of both hybrids. Importantly, the number of leaves and SPAD value were non-destructive and easy to investigate in response to water-deficit stress, suggesting that they may be useful indicators for screening drought-tolerant genetic resources. We detected more than 100 spots that were differentially accumulated under drought stress. Of these spots, a total of 21 protein spots (≥1.5-fold) from drought-exposed maize leaves were successfully analyzed by MALDI-TOF-TOF mass spectrometry. Functional annotation using Gene Ontology analysis revealed that most of the identified proteins were involved in carbohydrate metabolism, stress response fatty acid catabolism, photosynthesis, energy metabolism, and transport. The protein expression levels were increased in both Ilmichal and Gwangpyeongok, except for triosephosphate isomerase, fructose-bisphosphate aldolase, and an uncharacterized protein. The lactoylglutathione lyase delta (3,5)-delta (2,4)-dienoyl-CoA isomerase was overexpressed in Gwangpyeongok only. The results obtained from this study suggest that the drought-specific genes may be useful as molecular markers for screening drought-tolerant maize genotypes.

Expression of Cell Cycle Related Genes in HL60 Cells Undergoing Apoptosis by X-irradiation (HL60 세포주에서 방사선 조사에 의한 Apoptosis와 세포 주기 관련 유전자의 발현 변화)

  • Kim, Jin-Hee;Park, In-Kyu
    • Radiation Oncology Journal
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    • v.16 no.4
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    • pp.377-388
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    • 1998
  • Purpose : To evaluate changes in expression of cell cycle related genes during apoptosis induced in HL60 cells by X-irradiation to understand molecular biologic aspects in mechanism of radiation therapy. Material and Methods : HL-60 cell line (promyelocytic leukemia cell line) was grown in culture media and irradiated with 8 Gr by linear accelerator (6 MV X-ray). At various times after irradiation, ranging from 3 to 48 hours were analyzed apoptotic DNA fragmentation assay for apoptosis and by western blot analysis and semi-quantitative RT-PCR for expression of cell cycle related genes (cyclin A, cyclin B, cyclin C, cyclin Dl, cyclin E, cdc2, CDK2, CDK4, $p16^{INK4a}$, $p21^{WAF1}$, $p27^{KIP1}$, E2F, PCNA and Rb). Results : X-irradiation (8 Gy) induced apoptosis in HL-60 cell line. Cycline A protein increased after reaching its peak 48 h after radiation delivery and cyclin E, E2F, CDK2 and RB protein increased then decreased after radiation. Radiation induced up-regulation of the expression of E2F is due to mostly increase of Phosphorylated retinoblastoma proteins (ppRb). Cyclin Dl, PCNA, CDC2, CDK4 and $p16^{INK4a}$ protein underwent no significant change at any times after irradiation. There was not detected $p21^{WAF1}$ and $p27^{KIP1}$ protein. Cyclin A, B, C mRNA decreased immediately after radiation and then increased at 12 h after radiation. Cyclin Dl mRNA increased immediately and then decreased at 48 h after radiation. After radiation, cyclin E mRNA decreased with the lapse of time. CDK2 mRNA decreased at 3h and increased at eh after radiation. CDK4 mRNA rapidly increased at 6 to 12 h after radiation. There was no change of expression of $p16^{INK4a}$ and not detected in expressin of $p21^{WAF1}$ and $p27^{KIP1}$ mRNA. Conclusion : We suggest that entry into S phase may contribute to apoptosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of pRb protein are related with radiation induced auoptosis of HL60 cells and tosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of PRb protein are related with radiation induced apoptosis of HL60 cells and this may be associated with induction of E2F and cyclinE/CDK2. These results support that $p21^{WAF1}$ and $p27^{KIP1}$ are not related with radiation induced-apoptosis.

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Molecular Characterization and Toxin Profile of Bacillus cereus Strains Isolated from Ready-to-eat Foods (유통 중인 즉석·편의식품류에서 분리한 Bacillus cereus의 산생 Toxin 및 분자유전학적 특성 조사)

  • Kim, Tae Sun;Kim, Min Ji;Kang, Yu Mi;Oh, Geune;Choi, Su Yeon;Oh, Mu Sul;Yang, Yong Shik;Seo, Jung-Mi;Ryu, Mi-Geum;Kim, Eun-Sun;Ha, Dong-Ryong;Cho, Bae Sik
    • Korean Journal of Food Science and Technology
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    • v.46 no.3
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    • pp.334-340
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    • 2014
  • Toxin-producing Bacillus cereus is the causative agent of two different types of food poisoning: the emetic and the diarrheal types. This study was conducted to investigate the presence of enterotoxin and emetic toxin genes in 263 B. cereus isolated from 619 different ready-to-eat food items. Hemolytic enterotoxins hblA, hblC, and hblD were detected in 85.6, 41.1, and 76.8%, respectively, of the B. cereus isolates. About 67.0% (175/263) of the isolates presented all of three genes. Non-hemolytic enterotoxins nheA, nheB, and nheC were detected in 100, 97.0, and 68.4% of the isolates, respectively. Approximately 90.0% (236/263) of the isolates presented all of these three non-hemolytic enterotoxin genes. Emetic toxin gene, CER, was detected in 132 of 263 (50.2%) isolates. Computer-assisted cluster analysis of Rep-PCR profiles showed a high genetic diversity among the isolates. All B. cereus isolates from food samples tested in this study carried at least 6 of 10 toxin genes.

Isolation and Characterization of a Marine Bacterium, Pseudomonas sp. YJ-1 with Anti-Methicillin Resistant Staphylococcus aureus Activity (항 Methicillin Resistant Staphylococcus aureus 활성을 가지는 해양미생물 Pseudomonas sp. YJ-1의 분리와 특성)

  • Woo, Ye-Ju;Jeong, Seong-Yun
    • Korean Journal of Environmental Biology
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    • v.35 no.4
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    • pp.694-705
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    • 2017
  • The aim of this study was to isolate and identify marine bacterium with anti-methicillin-resistant Staphylococcus aureus (MRSA) activity, and to purify the anti-MRSA compound, as well as to determine its activity and synergistic effects. Among the marine bacteria isolated in this study, the YJ-1 isolate had the strongest anti-MRSA activity. The YJ-1 isolate was identified on the basis of its biochemical characteristics and an analysis of 16S rRNA gene sequences. The YJ-1 isolate showed over 99.2% homology with Pseudomonas stutzeri, and was designated as a Pseudomonas sp. YJ-1. The optimal culture conditions were $25^{\circ}C$ and initial pH 7.0. For the purification of the anti-MRSA compounds, the YJ-1 was cultured in Pa PES-II medium, and the culture filtrates were extracted by ethyl acetate, hexane, and 80% MeOH. The 80% MeOH fraction was separated by a $C_{18}$ ODS column, silica gel chromatography and a reverse phase HPLC, to yield three anti-MRSA agents, the MR1, MR2, and MR3 compounds. When the MR1 compound of $250{\mu}g\;mL^{-1}$ concentration was applied to the MRSA cells, over 95% of bacterial cells was killed within 48 hr. Compared with vancomycin and ampicillin, the MR1 compound showed significant anti-MRSA activity. In addition, the anti-MRSA activity was increased by dose and time dependent manners. Furthermore, the combination of an MR1 compound with vancomycin produced a more rapid decrease in the MRSA cells than did the MR1 compound alone. Taken together, our results suggest that the Pseudomonas sp. YJ-1 and its anti-MRSA compounds could be employed as a natural antibacterial agent in MRSA infections.

Anti-inflammatory Activities of Antimicrobial Peptide Locustacin Derived from Locusta migratoria in LPS-stimulated RAW264.7 Cells (풀무치 유래 항균 펩타이드 locustacin의 항염증 활성)

  • Choi, Ra-Yeong;Lee, Joon Ha;Seo, Minchul;Kim, In-Woo;Hwang, Jae-Sam;Kim, Mi-Ae
    • Journal of Life Science
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    • v.31 no.10
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    • pp.898-904
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    • 2021
  • Locusta migratoria is a widespread locust species in many parts of the world and is considered an alternative source for the production of protein for value-added ingredients. We previously identified putative antimicrobial peptides derived from L. migratoria through an in silico analysis of its transcriptome. However, its anti-inflammatory effect has not been studied. In this study, we investigated the anti-inflammatory activities of the antimicrobial peptide locustacin (KTHILSFFPSFLPLFLKK-NH2) derived from L. migratoria on lipopolysaccharide (LPS)-stimulated RAW264.7 macrophage cells. Locustacin (50, 100, and 200 ㎍/ml) significantly reduced the production of nitric oxide (NO) in LPS-stimulated macrophages without any cytotoxicity. Locustacin also inhibited the mRNA and protein expression of pro-inflammatory mediators, such as inducible NO synthase and cyclooxygenase-2, in contrast to the presence of LPS alone. Locustacin decreased the release of LPS-induced pro-inflammatory cytokines, including interleukin (IL)-6 and IL-1β, and their gene expression in a dose-dependent manner. Furthermore, locustacin (100 and/or 200 ㎍/ml) inhibited phosphorylation levels of extracellular signal regulated kinase, p38, and c-Jun N-terminal kinase. Locustacin also suppressed the degradation of inhibitory kappa B alpha, which was considered to be an inhibitor of nuclear factor kappa B (NF-κB). Collectively, these results demonstrate that locustacin can exert anti-inflammatory effects through the inhibition of mitogen-activated protein kinase (MAPK) phosphorylation, activation of NF-κB, and downstream inflammatory mediators in LPS-stimulated macrophage cells.