• Title/Summary/Keyword: gene gun

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Report of eight unrecorded Acetobacter species in Korea, discovered during the survey in 2018-2019

  • Heo, Jun;Won, Miyoung;Lee, Daseul;Han, Byeong-Hak;Hong, Seung-Beom;Kwon, Soon-Wo
    • Journal of Species Research
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    • v.11 no.3
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    • pp.155-161
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    • 2022
  • Acetic acid bacteria (AAB) convert ethanol to acetic acid through oxidation, and the fermentation pathway of AAB is important in the vinegar industry. The genus Acetobacter is the representative one of AAB, and several Korean traditional vinegars are produced using Acetobacter strains. Until now, four species in the genus Acetobacter were reported as native species in Korea. During the past two years, we isolated several AAB strains from fruits, flowers and fermented foods, and several AAB species unrecorded in Korea were found on the basis of 16S rRNA gene sequence analyses. In this study, we report eight Acetobacter species as native ones which are A. fabarum C10-3 (=KACC 21483) isolated from plumcot fruit (Naju-si), A. lovaniensis KDG-EC1 (=KACC 22697) isolated from diced radish kimchi (Naju-si), A. okinawensis GAM12-M2 (=KACC 22696) isolated from persimmon fruit (Sangju-si), A. orientalis FR32C4 (=KACC 22370) isolated from fruit of Cudrania tricuspidata (Jeonju-si), A. papaya FR35B3 (=KACC 22046) isolated from grape fruit (Yeongdong-gun), A. suratthaniensis GAM15-R2 (=KACC 22694) isolated from persimmon fruit(Gimje-si), A. syzygii C25-1 (=KACC 22048) isolated from peach fruit (Namwon-si) and A. thailandicus JDF1-M1 (=KACC 22693) isolated from plum fruit(Seoul).

Identifying the Anti-Cancer Effect of Indigo Naturalis in Small Cell Lung Cancer Based on Network Pharmacological Analysis (네트워크 약리학적 분석에 의한 소세포폐암에 대한 청대의 항암기전 연구)

  • Young Hoon, Kim;Woo Jin, Jeong;Gwang Hee, Jeong;Youn Sook, Kim;Won Gun, An
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.36 no.6
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    • pp.229-234
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    • 2022
  • Lung cancer is the leading cause of cancer-related deaths worldwide. Indigo Naturalis (IN) is a dark blue powder obtained by processing leaves or stems of indigo plants, its anticancer effects have been reported in several studies. However, the pharmacological mechanism of IN in small cell lung cancer (SCLC) is not elucidated. In this study, to investigate the anticancer efficacy of IN for SCLC, we presented potential active ingredients, SCLC-related targets, and pharmacological mechanisms of IN that are expected to have anticancer activity for SCLC using a network pharmacological analysis. The phytochemical compounds of IN have been collected through TCMSP, SymMap, or HPLC documents. The active ingredients of IN such as indirubin, indican, isatin, and tryptanthrin were selected through ADME parameters or literature investigations for each compound. Using the Compounds, Disease-Target associations Databases, 124 common targets of IN and SCLC were obtained. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) Pathway enrichment analysis was carried out. GO biological processes are associated with response to xenobiotic stimulus, positive regulation of protein phosphorylation, regulation of mitotic cell cycle, and regulation of apoptotic signaling pathway. KEGG disease pathways included Gastric cancer, Bladder cancer, SCLC, and Melanoma. The main anticancer targets of the IN for SCLC were analyzed in 14 targets, including BCL2, MYC, and TP53. In conclusion, the results of this study based on the network pharmacology of IN can provide important data for the effective prevention and treatment of SCLC.

Systems Pharmacological Approach to Identification of Schizonepeta teunifolia Extract via Active Ingredients Analysis and Cytotoxicity Effect on A549 Cell Lines (형개 추출물의 시스템 약리학적 분석과 비소세포폐암세포에 대한 증식 억제효과)

  • Ga Ram Yang;Ji Eun Choo;Youn Sook Kim;Won Gun Ahn
    • Korean Journal of Acupuncture
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    • v.41 no.1
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    • pp.7-15
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    • 2024
  • Objectives : This study aimed to predict the effectiveness and potential of Schizonepeta tenuifolia as an anticancer treatment for non-small cell lung cancer through network-based pharmacology and cellular experiment. Methods : To identify the major bioactive compounds in Schizonepeta tenuifolia, we used the Traditional Chinese Medicine Systems. The target genes for the cancer treatment were selected using the UniProt database and the networked using Cytoscape. We performed functional enrichment analysis based on the Gene Ontology Biological Process and Kyoto Encyclopedia of Genes and Genomes Pathways to predict the mechanisms. To investigate the effect of Schizonepeta tenuifolia on lung cancer cell growth, we treated A549 cells, a lung cancer cell line, with different concentrations of the drug and used the MTT assay for cell viability. Results : Research has shown that the most effective mechanism of active compounds from Schizonepeta tenuifolia is through the pathway of cancer. The results of the network pharmacology analysis indicate that Schizonepeta tenuifolia has potential medicinal value as an adjuvant in anticancer treatment. The concentration-dependent inhibition of cell viability was observed on A549 cells. Furthermore, synergistic anticancer activity with Doxorubicin was also observed. Conclusions : Through a network pharmacological approach, Schizonepeta tenuifolia was predicted to have potential as an anticancer agent, and its efficacy was experimentally demonstrated using A549 cells. These findings suggest that Schizonepeta tenuifolia is a promising candidate for future research.

Characterization and Evaluation of Melanocortin 4 Receptor (MC4R) Gene Effect on Pork Quality Traits in Pigs (돼지 Melanocortin 4 Receptor (MC4R) 유전자의 육질연관성 분석)

  • Roh, Jung-Gun;Kim, Sang-Wook;Choi, Jung-Suk;Choi, Yang-Il;Kim, Jong-Joo;Choi, Bong-Hwan;Kim, Tae-Hun;Kim, Kwan-Suk
    • Journal of Animal Science and Technology
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    • v.54 no.1
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    • pp.1-8
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    • 2012
  • This study aimed to investigate the single nucleotide polymorphisms (SNPs) of the porcine MC4R gene and validate the effect of the MC4R genotype for marker assisted selection (MAS). Six amplicons were produced to analyze the entire base sequences of the porcine MC4R gene and six SNPs were detected (c.-780C>G, c.-135C>T, c.175C>T-Leu59Leu, c.707A>G-Arg236His, c.892A>G-Asp298Asn, and c.*430A>T). Linkage disequilibrium (LD) of the six SNPs was analyzed by performing haploid analysis. There was a perfect linkage disequilibrium in c.-780C>G, c.-135C>T, c.175C>T-Leu59Leu, c.707A>G-Arg236His, and c.*430A>T. Only the c.892A>G (Asp298Asn) SNP showed a very low LD with an $r^2$ value of 0.028 and the D' value of 0.348. As a result, the two SNPs-c.707A>G (Arg236His) and c.892A>G (Asp298Asn)-were selected to extract the genotype frequencies from the 5 pig breeds by using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) genotype analysis method. The SNP frequency of c.707A>G (Arg236His) indicated the presence of the A (His) allele only in Yorkshire, while the G allele was fixed in the KNP, Landrace, Berkshire, and Duroc. Association analysis was carried out in 484 pigs with the c.707A>G (Arg236His) SNP and the meat quality traits of four different pig cross populations: a significant association was noted in crude fat, sirloin moisture, meat color, and the degree of red and yellow coloration. The frequency of the c.892A>G(Asp298Asn) SNP genotype varied among the breeds; while Duroc showed the highest frequency of the A (Asn) allele, KNP showed the highest frequency of the G (Asp) allele. Association analysis was carried out in 1126 pigs with the c.892A>G (Asp298Asn) SNP and the meat quality traits of four pig populations: a highly significant linkage was noted in the back-fat thickness (P<0.002). It was found that the back-fat thickness was higher in individuals with the AA genotype than in those with the AG or GG genotype. Thus, in this study, we verified that the c.892A>G (Asp298Asn) SNP in the pig MC4R gene has a sufficient effect as a gene marker for MAS in Korean pork industry.

Lack of the Association between Microsatellite Polymorphism in Toll-like Receptor 2 Gene and Development of COPD (Toll-like Receptor 2 유전자의 Microsatellite 유전자 다형성과 만성폐쇄성폐질환 발생과의 연관성 결여)

  • Lee, Hee Seok;Lee, Hye Won;Kim, Deog Kyeom;Ko, Dong Seok;Park, Gun Min;Hwang, Yong Il;Lee, Sang-Min;Yoo, Chul Gyu;Kim, Young Whan;Han, Sung Koo;Shim, Young-Soo;Yim, Jae-Joon
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.4
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    • pp.367-374
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    • 2005
  • Background : The fact that only 10-20% of chronic cigarette smokers develop chronic obstructive pulmonary disease (COPD) reflects the presence of genetic factors associated with the susceptibility to COPD. Recently, it was reported that the surfactant protein A increases the secretion of matrix metalloprotease 9, which degrades extracellular matrices of the lung, through a Toll-like receptor 2 (TLR2). In this context, possible role of TLR2 in the pathogenesis of COPD was postulated, and a functional dinucleotide repeat polymorphism in intron II of TLR2 was evaluated for any association with COPD. Method : Male patients with COPD and male smokers with a normal pulmonary function were enrolled in this study. The number of Guanine-Thymine repeats in intron II of the TLR2 gene were counted. Because the distributions of the repeats were trimodal, the alleles were classified into three subclasses, 12-16 repeats: short (S) alleles; 17-22 repeats: medium length (M) alleles; and 23-27 repeats: long (L) alleles. Result : 125 male patients with COPD and 144 age- and gender-matched blood donors with a normal lung function were enrolled. There were no differences in the distribution of each allele subclass (S, M and L) between the COPD and control group (p=0.75). The frequencies of the genotypes with and without each allele subclass in the COPD and control group were similar. Conclusion : A microsatellite polymorphism in intron II of TLR2 gene was not associated with the development of COPD in Koreans.

Purification, Characterization, and Cloning of Fibrinolytic Metalloprotease from Pleurotus ostreatus Mycelia

  • Shen, Ming-Hua;Kim, Jae-Sung;Sapkota, Kumar;Park, Se-Eun;Choi, Bong-Suk;Kim, Seung;Lee, Hyun-Hwa;Kim, Chun-Sung;Chun, Hong-Sung;Ryoo, Cheon-In;Kim, Sung-Jun
    • Journal of Microbiology and Biotechnology
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    • v.17 no.8
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    • pp.1271-1283
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    • 2007
  • A fibrinolytic protease (PoFE) was purified from the cultured mycelia of the edible oyster mushroom Pleurotus ostreatus, using a combination of various chromatographies. The purification protocol resulted in an 876-fold purification of the enzyme, with a final yield of 6.5%. The apparent molecular mass of the purified enzyme was estimated to be 32 kDa by SDS-PAGE, fibrin-zymography, and size exclusion using FPLC. The optimal reaction pH value and temperature were pH 6.5 and $35^{\circ}C$, respectively. PoFE effectively hydrolyzed fibrinogen, preferentially digesting the $A{\alpha}$-chain and the $B{\beta}$-chain over the ${\gamma}$-chain. Enzyme activity was enhanced by the addition of $Ca^{2+},\;Zn^{2+},\;and\;Mg^{2+}$ ions. Furthermore, PoFE activity was potently inhibited by EDTA, and it was found to exhibit a higher specificity for the chromogenic substrate S-2586 for chymotrypsin, indicating that the enzyme is a chymotrypsin-like metalloprotease. The first 19 amino acid residues of the N-terminal sequence were ALRKGGAAALNIYSVGFTS, which is extremely similar to the metalloprotease purified from the fruiting body of P. ostreatus. In addition, we cloned the PoFE protein, encoding gene, and its nucleotide sequence was determined. The cDNA of cloned PoFE is 867 nucleotides long and consists of an open reading frame encoding 288 amino acid residues. Its cDNA showed a high degree of homology with PoMEP from P. ostreatus fruiting body. The mycelia of P. ostreatus may thus represent a potential source of new therapeutic agents to treat thrombosis.

Phylogenetic Study of Genus Haliotis in Korea by Cytochrome c Oxidase Subunit 1 and RAPD Analysis (Cytochrome c oxidase subunit 1과 RAPD 분석에 의한 한국 전복속의 계통 연구)

  • Seo, Yong Bae;Kang, Sung Chul;Choi, Seong Seok;Lee, Jong Kyu;Jeong, Tae Hyug;Lim, Han Kyu;Kim, Gun-Do
    • Journal of Life Science
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    • v.26 no.4
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    • pp.406-413
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    • 2016
  • Abalones are gastropod mollusks belonging to the genus Haliotis. Pacific abalones are regarded as a very important marine gastropod mollusk in Korea, Japan, China, and also in food industries around the world. In Korea, 6 species of abalone have been reported to occur along the coasts: Haliotis discus hannai, Haliotis discus discus, Haliotis madaka, Haliotis gigantea, Haliotis diversicolor supertexta, and Haliotis diversicolor diversicolor. This study was performed to discriminate the genetic variances by the partial sequences of the mitochondrial cytochrome c oxidase subunit I (COI) genes and random amplified polymorphic DNA (RAPD) analysis against four species of Pacific abalone (H. discus hannai, H. discus, H. madaka, H. gigantea). COI gene is reasonably well conserved and has been sequenced in various invertebrate taxa. The RAPD analysis technique is a relatively simple and low cost method that allows differentiation of taxa without the need to know their genomes. In this study, we investigated the genetic diversity, phylogenetic relationships within each species. The COI and RAPD analysis were able to distinguish between H. gigantea and the other three species. However, these analysis methods were inadequate to distinguish between H. discus and H. madaka. These results are believed to be able to provide a basis data for future hybrid breeding research by defining the genetically closely related four species of abalone, which is to develop new hybrid abalone for export using hybrid breeding.

Plant Growth Promotion and Gibberellin A3 Production by Aspergillus flavus Y2H001 (Aspergillus flavus Y2H001의 식물생육촉진과 Gibberellin A3의 생산)

  • You, Young-Hyun;Park, Jong Myong;Kang, Sang-Mo;Park, Jong-Han;Lee, In-Jung;Kim, Jong-Guk
    • The Korean Journal of Mycology
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    • v.43 no.3
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    • pp.200-205
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    • 2015
  • Perilla frutescens var. japonica Hara was collected from farmland in Seongju-gun. Fifteen endophytic fungal strains with different colony morphologies were isolated from roots of P. frutescens. Waito-c rice seedlings were treated with the concentrated culture filtrates (CF) of endophytic fungi for observation of their plant growth-promoting activities. In the results, the CF of Y2H001 fungal strain promoted the growth of the waito-c rice seedlings. The phylogenetic tree of Y2H001 strain was analyzed by the combined sequences of the partial internal transcribed spacer region (ITS) and partial betatubulin gene. Molecular and morphological studies identified the Y2H001 strain as belonging to Aspergillus flavus. In gas chromatography mass spectrometry (GC/MS) analysis of the CF of Y2H001 strain, gibberellic acid (GA) was detected and quantified. Therefore, we describe Y2H001 strain as a new $GA_3$-producing A. flavus based on morphological, molecular characteristics and analysis of secondary metabolite.

Analysis of Varietal Differences in Pre-harvest Sprouting of Rice using RNA-Sequencing (RNA-Sequencing을 이용한 벼 품종간 수발아 차이 분석)

  • Choi, Myoung-Goo;Lee, Hyen-Seok;Hwang, Woon-Ha;Yang, Seo-Yeong;Lee, Yun-Ho;Lee, Chung-gun;Yun, Song Joong;Jeong, Jae-Hyeok
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.65 no.4
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    • pp.274-283
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    • 2020
  • Seed dormancy is an adaptive trait in which seeds do not germinate under unfavorable environmental conditions. Low dormancy seeds are easily germinated under optimal environmental conditions, and these characteristics greatly reduce the yield and quality of crops. In the present study, we compared the pre-harvest sprouting (PHS) rate of two cultivars, Joun and Jopyeong, using the Winkler scale after heading day and temperature of the test. The PHS rate increased as the Winkler scale after heading day increased from 700℃ to 1100℃ and the temperature of the test increased. In all conditions, the PHS rate of Jopyeong was higher than that of Joun. RNA-sequencing was used to analyze the cause of the high PHS rate. We analyzed the biological metabolic processes related to the abscisic acid (ABA) metabolite pathway using the KEGG mapper with selected differentially expressed genes in PHS seeds. We found that the expression of ABA biosynthesis genes (OsNCEDs) was down-regulated and that ABA catabolic genes (OsCYP707As) was up-regulated in PHS seeds. However, the quantitative real-time PCR results showed that Joun had a higher expression of OsNCEDs than that of Jopyeong, but OsCYP707As did not yield a significant result. Joun displayed higher ABA content than that of Jopyeong not only during ripeness time but also during PHS treatment. Taken together, we provided evidence that the ABA content remaining in the seed is important to the PHS rate, which is determined by the expression level of the ABA biosynthesis gene OsNCEDs.

Efficient Transduction with Recombinant Adenovirus in EBV-transformed B Lymphoblastoid Cell Lines

  • Kim, Hye-Jin;Cho, Hyun-Il;Han, Yoon-Hee;Park, Soo-Young;Kim, Dong-Wook;Lee, Dong-Gun;Kim, Jee-Hoon;Shin, Wan-Shik;Paik, Soon-Young;Kim, Chun-Choo;Hong, Young-Seon;Kim, Tai-Gyu
    • BMB Reports
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    • v.37 no.3
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    • pp.376-382
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    • 2004
  • The Epstein-Barr-transformed B lymphoblastoid cell lines, LCL, which express antigens, are potential antigen-presenting cells (APCs) for the induction of cytotoxic T lymphocytes in vitro. However, transfecting LCL with subsequent selection by antibiotics is notoriously difficult because the plating efficiencies of LCL are reported to be 1% or less. Therefore, this study investigated the optimal conditions for increasing the transduction efficiency of a recombinant adenovirus to LCL for use as a source of APCs. The transduction efficiencies were < 13% (SD $\pm$ 2.13) at a multiplicity of infection (MOI) of 100, while it was increased to 28% (SD $\pm$ 9.43) at an MOI of 1000. Moreover, its efficiencies to LCL that expressed the coxsackie adenovirus receptor were increased to 60% (SD $\pm$ 6.35) at an MOI of 1000, and were further increased to 70% (SD $\pm$ 4.56) when combined with the centrifugal method. The cationic liposome or anionic polymer had no effect on the transduction efficiency when compared to that of the centrifugal method. These results may be used as a convenient source of target cells for a CTL assay and/or autologous APCs for the induction of the in vitro CTL responses that are specific to viral and tumor antigens.