• Title/Summary/Keyword: gene screening

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Influence of the plant growth promoting Rhizobium panacihumi on aluminum resistance in Panax ginseng

  • Kang, Jong-Pyo;Huo, Yue;Yang, Dong-Uk;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.45 no.3
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    • pp.442-449
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    • 2021
  • Background: Panax ginseng is an important crop in Asian countries given its pharmaceutical uses. It is usually harvested after 4-6 years of cultivation. However, various abiotic stresses have led to its quality reduction. One of the stress causes is high content of heavy metal in ginseng cultivation area. Plant growth-promoting rhizobacteria (PGPR) can play a role in healthy growth of plants. It has been considered as a new trend for supporting the growth of many crops in heavy metal occupied areas, such as Aluminum (Al). Methods: In vitro screening of the plant growth promoting activities of five tested strains were detected. Surface-disinfected 2-year-old ginseng seedlings were dipping in Rhizobium panacihumi DCY116T suspensions for 15 min and cultured in pots for investigating Al resistance of P. ginseng. The harvesting was carried out 10 days after Al treatment. We then examined H2O2, proline, total soluble sugar, and total phenolic contents. We also checked the expressions of related genes (PgCAT, PgAPX, and PgP5CS) of reactive oxygen species scavenging response and pyrroline-5-carboxylate synthetase by reverse transcription polymerase chain reaction (RT-PCR) method. Results: Among five tested strains isolated from ginseng-cultivated soil, R. panacihumi DCY116T was chosen as the potential PGPR candidate for further study. Ginseng seedlings treated with R. panacihumi DCY116T produced higher biomass, proline, total phenolic, total soluble sugar contents, and related gene expressions but decreased H2O2 level than nonbacterized Al-stressed seedlings. Conclusion: R. panacihumi DCY116T can be used as potential PGPR and "plant strengthener" for future cultivation of ginseng or other crops/plants that are grown in regions with heavy metal exposure.

Screening of Bacterial Antagonists to Develop an Effective Cocktail against Erwinia amylovora

  • Choi, Dong Hyuk;Choi, Hyun Ju;Kim, Yeon Ju;Lim, Yeon-Jeong;Lee, Ingyeong;Park, Duck Hwan
    • Research in Plant Disease
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    • v.28 no.3
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    • pp.152-161
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    • 2022
  • Several types of chemical bactericides have been used to control fire blight. However, their excessive usage leads to environmental deterioration. Therefore, several researchers have analyzed antagonistic microorganisms as promising, effective, and safe biological control agents (BCAs). The primary aim of this study was to screen for potential antagonistic bacteria that suppress Erwinia amylovora. Among the 45 isolates studied, 5 strains showed the largest inhibition zone against E. amylovora. 16S rRNA gene sequencing identified them as Bacillus amyloliquefaciens (KPB 15), B. stratosphericus (KPB 21), B. altitudinis (KPB 25), B. safensis (KPB 31), and B. subtilis (KPB 39). KPB 25 and 31 reduced the lesion size of fire blight by 50% in immature apple fruits, and did not show antagonism against each other. Therefore, KPB 25 and 31 were selected to develop an antagonistic mixture against fire blight. Although the mixture with KPB 25 and 31 showed a slightly increased ability to reduce lesion size on immature fruits, they did not exhibit a synergistic effect in reducing E. amylovora population compared to each strain alone. Nevertheless, we have identified these two strains as useful and novel BCAs against fire blight with additional benefits safety and potential in developing a mixture without loss of their activity, owing to the absence of antagonism against each other.

A Rapid and Universal Direct PCR Method for Macrofungi

  • Park, Mi-Jeong;Lee, Hyorim;Ryoo, Rhim;Jang, Yeongseon;Ka, Kang-Hyeon
    • The Korean Journal of Mycology
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    • v.49 no.4
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    • pp.455-467
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    • 2021
  • Macrofungi are valuable resources as novel drug candidates, new biomaterials, and edible materials. Recently, genetic approaches pertaining to macrofungi have been continuously growing for their identification, molecular breeding, and genetic engineering. However, purification and amplification of fungal DNA is challenging because of the rigid cell wall and presence of PCR inhibitory metabolites. Here, we established a direct PCR method to provide a rapid and efficient method for PCR-grade macrofungal DNA preparation applicable to both conventional PCR and real-time PCR. We first optimized the procedure of lysis and PCR using the mycelia of Lentinula edodes, one of the most widely consumed macrofungal species. Lysates prepared by neutralizing with (NH4)2SO4 after heating the mycelia in a mixture of TE buffer and KOH at 65℃ for 10 min showed successful amplification in both conventional and real-time PCR. Moreover, the addition of bovine serum albumin to the PCR mixture enhanced the amplification in conventional PCR. Using this method, we successfully amplified not only internal transcribed spacer fragments but also low-copy genes ranging in length from 500 to 3,000 bp. Next, we applied this method to 62 different species (54 genera) of macrofungi, including edible mushrooms, such as Pleurotus ostreatus, and medicinal mushrooms such as Cordyceps militaris. It was found that our method is widely applicable to both ascomycetes and basidiomycetes. We expect that our method will contribute to accelerating PCR-based approaches, such as molecular identification, DNA marker typing, gene cloning, and transformant screening, in macrofungal studies.

Screening and functional validation of lipid metabolism-related lncRNA-46546 based on the transcriptome analysis of early embryonic muscle tissue in chicken

  • Ruonan, Chen;Kai, Liao;Herong, Liao;Li, Zhang;Haixuan, Zhao;Jie, Sun
    • Animal Bioscience
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    • v.36 no.2
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    • pp.175-190
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    • 2023
  • Objective: The study was conducted to screen differentially expressed long noncoding RNA (lncRNA) in chickens by high-throughput sequencing and explore its mechanism of action on intramuscular fat deposition. Methods: Herein, Rose crown and Cbb broiler chicken embryo breast and leg muscle lncRNA and mRNA expression profiles were constructed by RNA sequencing. A total of 96 and 42 differentially expressed lncRNAs were obtained in Rose crown vs Cobb broiler chicken breast and leg muscle, respectively. lncRNA-ENSGALT00000046546, with high interspecific variability and a potential regulatory role in lipid metabolism, and its predicted downstream target gene 1-acylglycerol-3-phosphate-O-acyltransferase 2 (AGPAT2), were selected for further study on the preadipocytes. Results: lncRNA-46546 overexpression in chicken preadipocyte 2 cells significantly increased (p<0.01) the expression levels of AGPAT2 and its downstream genes diacylglycerol acyltransferase 1 and diacylglycerol acyltransferase 2 and those of the fat metabolism-related genes peroxisome proliferator-activated receptor γ, CCAAT/enhancer binding protein α, fatty acid synthase, sterol regulatory element-binding transcription factor 1, and fatty acid binding protein 4. The lipid droplet concentration was higher in the overexpression group than in the control cells, and the triglyceride content in cells and medium was also significantly increased (p<0.01). Conclusion: This study preliminarily concludes that lncRNA-46546 may promote intramuscular fat deposition in chickens, laying a foundation for the study of lncRNAs in chicken early embryonic development and fat deposition.

Minor alleles in the FTO SNPs contributed to the increased risk of obesity among Korean adults: meta-analysis from nationwide big data-based studies

  • Oh Yoen Kim;Jihyun Park;Jounghee Lee;Cheongmin Sohn;Mi Ock Yoon;Myoungsook Lee
    • Nutrition Research and Practice
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    • v.17 no.1
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    • pp.62-72
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    • 2023
  • BACKGROUND/OBJECTIVES: Many studies have revealed an association between fat mass and the obesity-related gene (FTO) and obesity. On the other hand, no meta-analysis was conducted with data from only Koreans. Therefore, this study performed a meta-analysis using Korean data to provide evidence for the association between FTO single nucleotide polymorphisms (SNPs) and the risk of obesity among Korean adults. SUBJECT/METHODS: Meta-analysis was finally conducted with data extracted from seven datasets of four studies performed on Korean adults after the screening passed. Five kinds of FTO SNPs (rs9939609, rs7193144, rs9940128, rs8050136, and rs9926289) were included, and the relationship between FTO SNPs and body mass index (BMI) was investigated using linear regression with an additive model adjusted for covariants, such as age, sex, and area. RESULTS: The minor alleles of FTO SNPs were associated with increased BMI (odds ratio [OR], 1.31; 95% confidence interval [CI], 1.21-1.42). In sub-group analysis, FTO rs9939609 T>A was significantly associated with BMI (OR, 1.23; 95% CI, 1.06-1.42). The other FTO SNPs together were significantly associated with BMI (OR, 1.37; 95% CI, 1.25-1.49). The publication bias was not observed based on Egger's test. CONCLUSIONS: This meta-analysis showed that minor alleles in the FTO SNPs were significantly associated with an increased BMI among Korean adults. This meta-analysis is the first to demonstrate that minor alleles in the FTO SNPs contribute significantly to the increased risk of obesity among Korean adults using data from a Korean population.

Alkaline Protease Production from Bacillus gibsonii 6BS15-4 Using Dairy Effluent and Its Characterization as a Laundry Detergent Additive

  • Polson Mahakhan;Patapee Apiso;Kannika Srisunthorn;Kanit Vichitphan;Sukanda Vichitphan;Sukrita Punyauppa-path;Jutaporn Sawaengkaew
    • Journal of Microbiology and Biotechnology
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    • v.33 no.2
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    • pp.195-202
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    • 2023
  • Protease is a widely used enzyme particularly in the detergent industry. In this research, we aimed to isolate alkaline protease-producing bacteria for characterization as a laundry detergent additive. The screening of alkaline protease production was investigated on basal medium agar plus 1% skim milk at pH 11, with incubation at 30℃. The highest alkaline protease-producing bacterium was 6BS15-4 strain, identified as Bacillus gibsonii by 16S rRNA gene sequencing. While the optimum pH was 12.0, the strain was stable at pH range 7.0-12.0 when incubated at 45℃ for 60 min. The alkaline protease produced by B. gibsonii 6BS15-4 using dairy effluent was characterized. The optimum temperature was 60℃ and the enzyme was stable at 55℃ when incubated at pH 11.0 for 60 min. Metal ions K+, Mg2+, Cu2+, Na+, and Zn2+ exhibited a slightly stimulatory effect on enzyme activity. The enzyme retained over 80% of its activity in the presence of Ca2+, Ba2+, and Mn2+. Thiol reagent and ethylenediaminetetraacetic acid did not inhibit the enzyme activity, whereas phenylmethylsulfonyl fluoride significantly inhibited the protease activity. The alkaline protease from B. gibsonii 6BS15-4 demonstrated efficiency in blood stain removal and could therefore be used as a detergent additive, with potential for various other industrial applications.

Strategic Use of QTL Mapping to Improve the Palatability of Rice

  • Yoon-Hee Jang;Jae-Ryoung Park;Eun-Gyeong Kim;Kyung-Min Kim
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.286-286
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    • 2022
  • The properties of starch play an important role in determining the palatability of rice. In addition, the gelatinization temperature (GT) of rice starch is an important factor in determining the quality of rice because it is related to the cooking time and texture of rice. For the development of high-quality rice, it is important to understand the genetic basis of palatability-related traits, and QTL analysis is an effective method to explain the genetic basis of variation in complex traits. QTL mapping related to alkali digestion value (ADV) of brown and milled rice was performed using the 120 Cheongcheong/Nagdong double haploid (CNDH) line. As a result, 12 QTLs related to ADV were detected, and 20 candidate genes were selected from the RM588-RM1163 region of chromosome 6 through screening by gene function analysis. The comparison of the relative expression level of candidate genes showed that OsSS1q6 is highly expressed in CNDH lines with high ADV in both brown rice and milled rice. In addition, OsSS1q6 has high homology with starch synthase 1 protein, and interact with various starch biosynthesis-related proteins, such as GBSSII, SBE, and APL. Therefore, we suggest that OsSS1q6 identified through QTL mapping could be one of the various genes involved in the GT of rice by regulating starch biosynthesis. This study can be used as basic data for breeding high-quality rice and provides a new genetic resource that can increase the palatability of rice.

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Analysis of the Effectiveness of Garlic on Gastrointestinal motility disorders using a network pharmacological method (네트워크 약리학 방법을 이용한 위장관 운동성 장애 관련 마늘의 효능 분석)

  • Na Ri Choi;Byung Joo Kim
    • Herbal Formula Science
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    • v.31 no.4
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    • pp.245-252
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    • 2023
  • Objectives : The purpose of this study was to explore the compounds, targets and related diseases of garlic by the approaches of network pharmacology and bioinformatics in traditional chinese medicine. Methods : We investigated components and their target molecules of garlic using SymMap and TCMSP and they were compared with analysis platform. Results : 56 potential compounds were identified in garlic, 26 of which contained target information, and it was found that these 26 compounds and 154 targets interact with each other through a combination of 243 compounds. In addition, Apigenin was linked to the most targeted gene (78) in 26 compounds, followed by Kaempferol (61 genes), Nicotic Acid (14 genes), Geraniol (11 genes), Eee (10 genes), and Sobrol A (9 genes). Among 56 potential compounds, three compounds (Kaempferol, Dipterocarpol, and N-Methyl cytisine) corresponded to the active compound by screening criterion Absorption, Distribution, Metabolism, Excretion (ADME). In addition, 12 compounds in 56 potential compounds were associated with gastrointestinal (GI) motility disorder. Among them, Kaempferol was a compound that met the ADME parameters and the rest were potential compounds that did not meet. Also, Kaempferol was closely related to GI motility disorder, indicating that this Kaempferol could be a candidate for potential medical efficacy. Conclusions : It shows the relationship between the compound of garlic, an herbal supplement, and the biological process associated with GI motility disorder. These results are thought to help develop strategies for treating GI motility disorders.

A comparison study of pathological features and drug efficacy between Drosophila models of C9orf72 ALS/FTD

  • Davin Lee;Hae Chan Jeong;Seung Yeol Kim;Jin Yong Chung;Seok Hwan Cho;Kyoung Ah Kim;Jae Ho Cho;Byung Su Ko;In Jun Cha;Chang Geon Chung;Eun Seon Kim;Sung Bae Lee
    • Molecules and Cells
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    • v.47 no.1
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    • pp.100005.1-100005.15
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    • 2024
  • Amyotrophic lateral sclerosis is a devastating neurodegenerative disease with a complex genetic basis, presenting both in familial and sporadic forms. The hexanucleotide (G4C2) repeat expansion in the C9orf72 gene, which triggers distinct pathogenic mechanisms, has been identified as a major contributor to familial and sporadic Amyotrophic lateral sclerosis cases. Animal models have proven pivotal in understanding these mechanisms; however, discrepancies between models due to variable transgene sequence, expression levels, and toxicity profiles complicate the translation of findings. Herein, we provide a systematic comparison of 7 publicly available Drosophila transgenes modeling the G4C2 expansion under uniform conditions, evaluating variations in their toxicity profiles. Further, we tested 3 previously characterized disease-modifying drugs in selected lines to uncover discrepancies among the tested strains. Our study not only deepens our understanding of the C9orf72 G4C2 mutations but also presents a framework for comparing constructs with minute structural differences. This work may be used to inform experimental designs to better model disease mechanisms and help guide the development of targeted interventions for neurodegenerative diseases, thus bridging the gap between model-based research and therapeutic application.

Lupus Heart Disease Modeling with Combination of Induced Pluripotent Stem Cell-Derived Cardiomyocytes and Lupus Patient Serum

  • Narae Park;Yeri Alice Rim;Hyerin Jung;Yoojun Nam;Ji Hyeon Ju
    • International Journal of Stem Cells
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    • v.15 no.3
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    • pp.233-246
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    • 2022
  • Background and Objectives: Systemic lupus erythematosus (SLE) is a chronic autoimmune disease mainly affecting young women of childbearing age. SLE affects the skin, joints, muscles, kidneys, lungs, and heart. Cardiovascular complications are common causes of death in patients with SLE. However, the complexity of the cardiovascular system and the rarity of SLE make it difficult to investigate these morbidities. Patient-derived induced pluripotent stem cells (iPSCs) serve as a novel tool for drug screening and pathophysiological studies in the absence of patient samples. Methods and Results: We differentiated CMs from HC- and SLE-iPSCs using 2D culture platforms. SLE-CMs showed decreased proliferation and increased levels of fibrosis and hypertrophy marker expression; however, HC-and SLE-monolayer CMs reacted differently to SLE serum treatment. HC-iPSCs were also differentiated into CMs using 3D spheroid culture and anti-Ro autoantibody was treated along with SLE serum. 3D-HC-CMs generated more mature CMs compared to the CMs generated using 2D culture. The treatment of anti-Ro autoantibody rapidly increased the gene expression of fibrosis, hypertrophy, and apoptosis markers, and altered the calcium signaling in the CMs. Conclusions: iPSC derived cardiomyocytes with patient-derived serum, and anti-Ro antibody treatment could serve in effective autoimmune disease modeling including SLE. We believe that the present study might briefly provide possibilities on the application of a combination of patient-derived materials and iPSCs in disease modeling of autoimmune diseases.