• Title/Summary/Keyword: RNA 안정성

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Overexpression of Termostable Bacillus sp. in Recombinant E.coli (재조합 E.coli에서 고온성 Bacillus 균주의 과발현에 관한 연구)

  • 서화정;이인선
    • Journal of Food Hygiene and Safety
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    • v.15 no.1
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    • pp.51-54
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    • 2000
  • In the 5'-flanking region of the D-AAT, AspAT and AlaDH gene, I found three or two pairs of sequences(designated as Pl, P2, P3) which show significant similarity to the E.coli consensus sequences of -35 and -10 for promoters. The spacing between -35 and -10 is 16 to 18bp in all the three putative promoters Pl, P2 and P3 which is in good agreement with the preferred spacer length in E.coli and in B.subtilis. Therefore, the putative promoters may also function to increase the efficiency of transcriptional initiation. The most stable, double-helical“Shine-Dalgarno”pairing is formed with a free energy change(ΔG) of -13.0 kcal/mol, -9.6 kcal/mol, -15.8 kcal/mol.

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Isolation and characterization of a Bacillus spp. for manufacturing the feed additives in livestock (가축의 보조사료 개발을 위한 Bacillus spp.의 분리 및 특성)

  • Park, Hae Suk;Jo, Seung Wha;Yim, Eun Jung;Kim, Yun Sun;Moon, Sung Hyun;Cho, Ho Seong;Kim, Hyun-Young;Cho, Yong Sik;Cho, Sung Ho
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.419-426
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    • 2015
  • The aims of this study were to isolate spore-forming Bacillus strains that exhibit high digestibility and anti-pathogenic bacteria toward feed for calves. Total 136 spore-forming strains were isolated from finished feeds and their ingredients. Among them, 93 strains were identified as Bacillus species when analyzed by 16S rRNA sequencing. For industrial use, three strains named as Bacillus licheniformis SHS14, B. subtilis LCB7, B. amyloliquefaciens LCB10 were selected after evaluating the industrial standards that are related with heat and acid resistance, enzyme activities, and anti-pathogenic activities against Samonella dublin ATCC15480 and E. coli K99. After each culture, 3 selected strains were mixed together at 1:1:1 (v/v/v) ratio and then prepared as the mixed starter culture for feeding. The changes in microbial community were analyzed via 16S rRNA metagenomics. The initial community ratio among three strains was maintained even after manufacturing into final products. Also, in vitro, enzymatic and anti-pathogenic activities were almost same as those when cultured in single culture, and results of anti-pathogenic activities conducted with calves showed 90% activities against lincomycin, which would be indicative of a promising feed starter.

Isolation of a Bacillus licheniformis DK42 Producing Cellulase and Xylanase, and Properties of the Enzymes (Cellulase 및 Xylanase를 분비하는 Bacillus licheniformis DK42의 분리 및 효소 특성)

  • Kim, Min Jeong;Lim, Soo Jin;Kang, Dae-Kyung
    • Journal of Animal Science and Technology
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    • v.50 no.3
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    • pp.429-436
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    • 2008
  • A bacterium producing cold-active cellulase and xylanase was isolated from pig feces. The isolate, DK42 strain, was found to be the Gram-positive, non-motile, catalase-positive, and spore-forming stain. Under an electron microscope, the cells were observed to be rod-shaped. The isolate was identified as Bacillus licheniformis DK42 on the basis of morphological and biochemical properties as well as 16S rRNA gene sequences. The characterization of crude cellulase and xylanase from B. licheniformis DK42 was investigated. Cellulase exhibited an optimum temperature and pH at 45℃ and 6.0, whereas xylanase exhibited an optimum temperature and pH at 55℃ and 6.0. Especially cellulase maintained approx. 50% of its maximum activity even at 10℃, indicating that it is cold-active. Both cellulase and xylanase were stable after 2hr at 35℃, whereas they lost their activities after 30min at 65℃.

Investigation of Variation in Bacterial Community Structure in Endangered Korean Fir Tree by Habitats (멸종위기종 구상나무 서식지별 세균 군집 구조 변이 조사)

  • Young Min Ko;Geun-Hye Gang;Dae Ho Jung;Youn-Sig Kwak
    • Research in Plant Disease
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    • v.30 no.2
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    • pp.165-175
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    • 2024
  • The Korean fir tree (Abies koreana), an endemic species of South Korea, is experiencing a severe decline in population due to climate change. Studies on the conservation of Korean fir have been extensive, yet research regarding its correlation with rhizosphere bacterial communities remains scarce, warranting further investigation. In this study, metagenome amplicon sequencing targeting the 16S rRNA V4 region was conducted to examine the presence of specific bacterial communities in Korean fir and to investigate potential differences based on habitat types (rhizosphere of native or cultivated trees, soil of dead trees, and bulk soil) and seasonal variations (April, June, September, November). Here we show that although we could not identify specific taxa highly specifically with Korean fir, the rhizosphere bacterial community in native trees exhibited less variability in response to seasonal changes compared to that in bulk soils. Suggesting the establishment of relatively stable bacterial populations around the Korean fir natural habitat. Further research on other types of rhizosphere and/or microbes is necessary to investigate the distinct relationship of Korean fir with microbial communities.

A novel anti-adhesion agent using DNA aptamers for Streptococcus mutans (DNA 앱타머를 이용한 Streptococcus mutans의 부착 억제)

  • Park, Byung-Ju;Ohk, Seung-Ho
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.3
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    • pp.382-388
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    • 2018
  • In this study, the SELEX method was used to screen for and select aptamers with high selectivity and affinity for Streptococcus mutans, which is the causative agent of dental caries. Aptamers are single stranded oligonucleotides of DNA or RNA with high selectivity and affinity for target molecules because of their specific three-dimensional structures that are used to collect biometric information. When compared to antibodies in vitro, aptamers are more advantageous because of their ease of use in the screening process, low cost, chemical stability, and lack of restrictions on the target molecules. We sorted DNA aptamers, which contain 44 bp or 38 bp primer sequences in 5' and 3', 39 bp random sequences in the middle.We then analyzed the character and affinity to S. mutans. Aptamers of specific oligonucleotides that combine with S. mutans were selected and these results are selectively fused to S. mutans. The results confirmed that DNA aptamers can be used for rapid diagnosis and treatment of dental caries caused by bacteria of the oral cavity, namely S. mutans.

Bacillus polyfermenticus CJ9, Isolated from Meju, Showing Antifungal and Antibacterial Activities (메주로부터 분리한 항진균 및 항세균 활성의 Bacillus polyfermenticus CJ9)

  • Jung, Ji-Hye;Chang, Hae-Choon
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.340-349
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    • 2009
  • A CJ9 bacterial strain, which showed antifungal and antibacterial activities, was isolated from meju and identified as Bacillus polyfermenticus based on Gram staining, biochemical properties, as well as its 16S rRNA sequence. B. polyfermenticus CJ9 showed the antimicrobial activity against the various pathogenic molds, yeasts, and bacteria. The antibacterial activity was stable in the pH 5.0~9.0, but the activity was lost at $37^{\circ}C$ for 24 hr. The antifungal activity was stable in the pH range of 3.0~9.0 and reduced at $121^{\circ}C$ for 15 min, but antifungal activity was not completely destroyed. The antibacterial activity was completely inactivated by proteinase K, protease, trypsin, and $\alpha$-chymotrypsin. The antifungal activity was also completely inactivated by protease and $\alpha$-chymotrypsin, and reduced its activity by proteinase which indicated that the antifungal and antibacterial compounds have proteineous nature. The apparent molecular mass of the partially purified antifungal compound, as indicated by using the direct detection method in Tricine-SDS-PAGE, was approximately 1.4 kDa. The molecular mass of the antibacterial compound could not be determined because of its heat-liable characteristic.

Molecular Cloning and Identification of Novel Genes, Gomsin, Characteristically Expressed in Snailfish, Liparis tanakae (꼼치에서 특징적으로 발현되는 새로운 유전자 곰신의 분리 및 동정)

  • 송인선;이석근;손진기
    • Development and Reproduction
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    • v.6 no.1
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    • pp.7-16
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    • 2002
  • In order to obtain the specific genes of snailfish a subtracted cDNA library was constructed, and analysed by sequencing and GenBank search. Among them C90-171 clone was turned out to be genes showing low homology and nonredundant genes. This novel clone was named Gomsin(C90-171). Gomsin was shown to be intensely expressed in the epithelial cells, some mesenchymal cells, and sheaths of muscle bundles in the result of immunohistochemistry. In the cross reaction assay of Gomsin antibody against various human tissues, the Gomsin was strongly expressed in the ductal and acinar cells of salivary glands, which was similar to the expression patterns of proline-rich proteins(PRPs) of human. The antibody raised against the Gomsin was clearly cross-reacted with human salivary PRPs and also recombinant proteins of human PRPs in the Western blot and immunoprecipitation analysis. Contrast to the salivary PRPs, the Gomsin was not easily degraded in the mixed saliva, but rapidly attacked on the cultured keratocytes in vitro. The simulated protein structure of Gomsin was similar to the whorled pattern of PRPs, even though the amino acid sequence of Gomsin was quite different from those of PRPs. These data suggest that the Gomsin is a characteristic matrix protein in the skin and body of snailfish, which is also utilized for the tissue protection in the similar way to the PRPs of human muco-secretory organs.

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Role of PKR and EGR-1 in Induction of Interleukin-S by Type B Trichothecene Mycotoxin Deoxynivalenol in the Human Intestinal Epithelial Cells (B형 트리코테센 곰팡이 독소 데옥시니발레놀에 의한 인체 장관 상피세포 염증성 인터루킨 8유도에서의 PKR과 EGR-1의 상호 역할 규명)

  • Park, Seong-Hwan;Yang, Hyun;Choi, Hye-Jin;Park, Yeong-Min;Ahn, Soon-Cheol;Kim, Kwan-Hoi;Lee, Soo-Hyung;Ahn, Jung-Hoon;Chung, Duk-Hwa;Moon, Yu-Seok
    • Journal of Life Science
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    • v.19 no.7
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    • pp.949-955
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    • 2009
  • Mucosal epithelia sense external stress signals and transmit them to the intracellular cascade responses. Ribotoxic stress-producing chemicals such as deoxynivalenol (DON) or other trichothecene mycotoxins have been linked with gastrointestinal inflammatory diseases by Fusarium-contamination. The purpose of this study was to test the hypothesis that DON evokes the epithelial sentinel signals of RNA-dependent protein kinase (PKR) and early growth response gene 1 (EGR-1), which together contribute to the pro-inflammatory cytokine interleukin 8 (IL-8) in human intestinal epithelial cells. PKR suppression by the dominant negative PKR expression attenuated DON-stimulated interleukin-8 production. Moreover, 1L-8 transcriptional activation by DON was also reduced by PKR inhibition in the human intestinal epithelial cells. Treatment with the PKR inhibitor also suppressed EGR-1 promoter activity, mRNA and protein induction, although mitogen-activated protein (MAP) kinases such as extracellular signal-regulated protein kinases (ERK) 1/2, p38, c-Jun N-terminal Kinase (INK) were little affected or even enhanced in presence of a PKR inhibitor. These patterns were also compared in the EGR-1-suppressed cells, which showed much more suppressed production of 1L-8. All things taken into consideration, DON-activated sentinel signals of EGR-1 via PKR mediated interleukin-8 production in human intestinal epithelial cells, which provide insight into the possible general mechanism associated with mucosal inflammation as an intestinal toxic insult by ribotoxic trichothecene mycotoxins.

Isolation and Characterization of 𝛽-Glucosidase-Producing Yeast, Rhodotorula sp. GYP-1 (𝛽-Glucosidase 생성 효모 Rhodotorula sp. GYP-1의 분리 및 특성)

  • Hyun-Soo Roh;Min-Young Kwon;Sol-Bi Kim;Jae-Eun Cho;Song-Ih Han
    • Journal of the Korean Applied Science and Technology
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    • v.40 no.5
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    • pp.1126-1135
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    • 2023
  • Nine microbial strains were isolated from the byproduct of ginseng processing and field of ginseng cultivation. Two strains among them were confirmed. Phylogenetic analysis of these 𝛽-Glucosidase strains confirmed that strain GYP-1 belongs to the Rhodotorula and strain GYP-3-3 belong to genus Brachybacterium. Rhodotorula sp. GYP-1 was finally selected due to its high biomass production. The 𝛽-Glucosidase activity of Rhodotorula sp. GPY-1 was assessed at 30 ℃, and Higher than 70% of the enzyme activity was maintained at the temperature range of 20-40℃. Although the optimum pH for the highest enzyme activity was pH 5.0, the enzyme was stable throughout the pH range of 5.0-8.0. In addition, Rhodotorula sp. demonstrated antifungal activity against the ginseng root rot disease caused by Botrytis.

Novel Real Time PCR Method for Detection of Plasmodium vivax (새로운 Real Time PCR 방법을 통한 Malaria(Plasmodium vivax)의 검출)

  • Ki, Yeon-Ah;Kim, So-Youn
    • Microbiology and Biotechnology Letters
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    • v.33 no.2
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    • pp.148-153
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    • 2005
  • Malaria is a re-emerging infectious disease that is spreading to areas where it had been eradicated, such as Eastern Europe and Central Asia. To avoid the mortality from malaria, early detection of the parasite is a very important issue. The peripheral blood smear has been the gold standard method for the diagnosis of malaria infection. Recently, several other methods have been introduced for quantitative detection of malaria parasites. Real time PCR that employs fluorescent labels to enable the continuous monitoring of PCR product formation throughout the reaction has recently been used to detect several human malaria parasites. 18S rRNA sequences from malaria parasites have been amplified using Taqman real time PCR assay. Here, a SYBR Green-based real time quantitative PCR assay for the detection of malaria parasite-especially, Plasmodium vivax - was applied for the evaluation of 26 blood samples from Korean malaria patients. Even though SYBR Green-based real time PCR is easier and cheaper than Taqman-based assay, SYBR Green-based assay cannot be used because 18S rRNA cannot be specifically amplified using 1 primer set. Therefore, we used DBP gene sequences from Plasmodium vivax, which is specific for the SYBR Green based assays. We amplified the DBP gene from the 26 blood samples of malaria patients using SYBR Green based assay and obtained the copy numbers of DBP genes for each sample. Also, we selected optimal reference gene between ACTB and B2M using real time assay to get the stable genes regardless of Malaria titer. Using selected ACTB reference genes, we successfully converted the copy numbers from samples into titer, ${\sharp}$ of parasites per microliter. Using the resultant titer from DBP based SYBER Green assay with ACTB reference gene, we compared the results from our study with the titer from Taqman-based assay. We found that our results showed identical tendency with the results of 18S rRNA Taqman assay, especially in lower titer range. Thus, our DBP gene-utilized real time assay can detect Plasmodium vivax in Korean patient group semi-quantitatively and easily.