• Title/Summary/Keyword: AmA1 gene

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Isolation and Identification of Oceanisphaera sp. JJM57 from Marine Red Algae Laurencia sp. (Ceramiales: Rhodomelaceae) (해양 홍조류 Laurencia sp. (Ceramiales: Rhodomelaceae)에서 분리한 Oceanisphaera sp. JJM57의 분리 및 동정)

  • Kim, Man-Chul;Dharaneedharan, S.;Moon, Young-Gun;Kim, Dong-Hwi;Son, Hong-Joo;Heo, Moon-Soo
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.58-63
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    • 2013
  • A taxonomic study was carried out to assess the phylogenetic characteristics of isolate JJM57 from marine red algae Laurencia sp. collected from intertidal zone in Jeju Island, South Korea. Comparative analysis of 16S rRNA gene sequence shows that this isolate belongs to the genus Oceanisphaera. It shows 98.02% and 97.7% sequence similarity with Oceanisphera litoralis DSM $15406^T$ and Oceanisphera donghaensis KCTC $12522^T$, respectively. Strain JJM57 is a Gram-negative, aerobic, moderately halophilic bacterium able to grow in different NaCl concentration ranges from 0.5 to 8.0% and at varying temperatures from 4 to $37^{\circ}C$. Sharing some of the physiological and biochemical properties with O. litoralis and O. donghaensis, JJM57 strain differs in the utilization of ethanol, proline, and alanine. The G+C contents of the strain JJM57 is 61.94 mol% and it is rich in $C_{16:1}$ ${\omega}7c$ and/or iso-$C_{15:0}$ 2-OH, $C_{16:0}$, and $C_{18:1}$ ${\omega}7c$ fatty acids. The DNA-DNA relatedness data separates the strain JJM57 from other species such as O. litoralis and O. donghaensis. On the basis of these polyphasic evidences, present study proposed that strain JJM57 (=KCTC 22371 =AM983543 =CCUG 60764) represents a novel bacterial species of Oceanisphaera.

Isolation and Characterization of Six Microorganisms from the Digestive Tract of the Cricket Gryllus bimaculatus (쌍별귀뚜라미(Gryllus bimaculatus) 소화기관에서 분리한 6종류의 특성규명)

  • Kwon, Kisang;Lee, Eun Ryeong;Yoo, Bo-Kyung;Ko, Young Hwa;Shin, Hyojung;Choi, Ji-Young;Kwon, O-Yu
    • Journal of Life Science
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    • v.27 no.9
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    • pp.1040-1046
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    • 2017
  • We describe the isolation and characterization of six different intestinal microorganisms from the digestive tract of the cricket Gryllus bimaculatus. Based on 16S rRNA gene sequences, we obtained six isolates belonging to four different genera: Staphylococcus, Bacillus, Citrobacter, and Proteus. All the isolates were resistant to ampicillin. Ampicillin is an irreversible inhibitor of the enzymeetranspeptidase, which is needed to make bacterial cell walls. None of the isolates were resistant to kanamycin, which binds to the 30S subunit of the bacterial ribosome and then inhibits total protein synthesis. Gram staining was conducted, in addition to morphological classification under a microscope. Four grampositive isolates and two gram-negative isolates were detected. The gram-positive isolates were GL1 (round shaped, 2 am in diameter), GL2 (rod shaped, $2.5{\mu}m$ in length), GL3 (rod shaped, $2{\mu}m$ in length), and GL4 (round shaped, $1.5{\mu}m$ in diameter). The gram-negative isolates were GL5 (rod shaped, $2{\mu}m$ in length) and GL6 (rod-shaped, $2.5{\mu}m$ in length). Notably, two of the isolates, GL2 and GL4, secreted specific extracellular proteins. These were determined by MALDI-TOF-MS spectral analysis to be a 87 kDa collagenase, 56 kDa hypothetical protein, and 200 kDa hypothetical protein. The six isolates in this study could be used for various biotechnological applications and pest management, both in the field and in greenhouse systems. In addition, it would be interesting to determine the relationship between these isolates and their host.

Assessment of the Specificity of A Hybridization of Surfactant Protein A by Addition of Non-specific Rat Spleen RNA (Surfactant Protein A mRNA을 이용한 유전자 재결합 반응에서 비특이성 RNA의 첨가에 의한 특이성 검정)

  • Kim, Byeong Cheol;Kim, Mi Ok;Kim, Tae-Hyung;Sohn, Jang Won;Yoon, Ho Joo;Shin, Dong Ho;Park, Sung Soo
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.4
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    • pp.393-404
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    • 2004
  • Background : Nucleic acid hybridization has become an essential technique in the development of our understanding of gene structure and function. The quantitative analysis of hybridization has been used in the measurement of genome complexity and gene copy number. The filter hybridization assay is rapid, sensitive and can be used to measure RNAs complementary to any cloned DNA sequence. Methods : The authors assessed the accuracy, linearity, correlation coefficient and specificity of the hybridization depending on the added dose(0, 1, 5, and $10{\mu}g$) of non-specific rat spleen RNA to hybridization of surfactant protein A mRNA. Filter hybridization assays were used to obtain the equation of standard curve and thereby to quantitate the mRNA quantitation. Results : 1. Standard curve equation of filter hybridization assay between counts per minute (X) and spleen RNA input (Y) was Y=0.13X-19.35. Correlation coefficient was 0.98. 2. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) was Y=0.00066X-0.046. Correlation coefficient was 0.99. 3. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $1{\mu}g$ spleen RNA was Y=0.00056X-0.051. Correlation coefficient was 0.99. 4. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $5{\mu}g$ spleen RNA was Y=0.00065X-0.088. Correlation coefficient was 0.99. 5. Standard curve equation of filter hybridization assay between counts per minute (X) and surfactant protein A mRNA transcript input (Y) after the addition of $10{\mu}g$ spleen RNA was Y=0.00051X-0.10. Correlation coefficient was 0.99. Conclusions : Comparison of cpm/filter in a linear range allowed accurate and reproducible estimation of surfactant protein A mRNA copy number irrespective of the addition dosage of non-specific rat spleen RNA over the range $0-10{\mu}g$.

IMMUNOHISTOCHEMICAL STUDY ON EXPRESSION OF APOPTOSIS RELATED PROTEINS IN DENTIGEROUS CYST AND AMELOBLASTOMA (함치성 낭종 및 법랑아세포종에 있어서 Apoptosis 관련 단백 발현에 관한 면역조직화학적 연구)

  • Choi, Jin-Young
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.22 no.1
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    • pp.15-21
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    • 2000
  • Ameloblasotma is slowly growing, locally invasive neoplasm with a potentially destructive behavior. The epithelium of ameloblastoma is thought to have an intrinsic growth potential and has been shown to present a higher rate of proliferation as compared to odontogenic cysts with low local recurrence rate. The molecular mechanisms that regulate the cell growth and invasion of ameloblastoma cells are unknown. Bcl-2 protein, which prevent apoptosis, is expressed in immortalized ameloblastoma cell line(AM-1)(Harada et al 1998). Expression of bcl-2 protein occurs in tooth germs, whose epithelial component may act as the histogenic precursor of ameloblastoma. Bax is considered as a main effector of apoptosis. Bax forms homodimers and also heterodimers with bcl-2. p53 tumor supressor gene participates not only in cell proliferation control but also in induction of apoptosis. The objective of the present study was to evaluate the apoptosis related protein expression in odontogenic cyst and ameloblastoma. A total of 10 dentigerous cysts and 16 ameloblastomas were used in the present study. Dentigerous cyst showed negative or slight positive for p53 and bcl-2 but strongly positive for bax, ameloblastoma, on the other hand, strongly positive for p53 and bcl-2 but weekly positive for bax. Bcl-2 was expressed for ameloblastoma mainly in outer layer or whole layer of epithelium and for dentigerous cyst mainly in basal layer. The difference in expression of apoptosis related protein in dentigerous cyst and ameloblastoma might explain the peculiar aggressive growth pattern of ameloblastoma.

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Introduction and Expression of PAP gene using Agrobacterium in Scrophularia buergeriana Miquel (Agrobacterium을 이용한 PAP 유전자의 현삼으로 도입 및 형질발현)

  • Yu, Chang-Yeon;Seong, Eun-Soo;Lim, Jung-Dae;Huang, Shan-Ai;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.2
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    • pp.156-165
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    • 2001
  • Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

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Postsurgical Wound Infection Caused by Mycobacterium conceptionense Identified by Sequencing of 16S rRNA, hsp65, and rpoB Genes in an Immunocompetent Patient (16S rRNA, hsp65, 및 rpoB 염기순서분석으로 동정한 Mycobacterium conceptionense에 의한 면역능이 정상인 환자에서 발생한 수술후 창상감염)

  • Lee, Ja Young;Kim, Si Hyun;Shin, Jeong Hwan;Lee, Hyun-Kyung;Lee, Young Min;Song, Sae Am;Bae, Il Kwon;Kim, Chang-Ki;Jun, Kyung Ran;Kim, Hye Ran;Lee, Jeong Nyeo;Chang, Chulhun L.
    • Annals of Clinical Microbiology
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    • v.17 no.1
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    • pp.23-27
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    • 2014
  • Rapidly growing mycobacteria are ubiquitous in the environment and are increasingly being recognized as opportunistic pathogens. Recently, a new species, Mycobacteium conceptionense, has been validated from the Mycobacterium fortuitum third biovariant complex by molecular analysis. However, there are few reports, and postsurgical wound infection by this species is rare. We report a case of postsurgical wound infection caused by M. conceptionense in an immunocompetent patient that was identified by a sequencing analysis of 16S rRNA, hps65, and rpoB genes.

Effect of Astragalus Membranaceus Extract against Improvement of Myelosuppression and Quality of Life in 5-Fluorouracil Treated Mice (황기 추출물이 5-Fluorouracil을 투여한 생쥐의 골수억제 및 삶의 질에 미치는 영향)

  • Kwon, Chang-Hyun;Yoo, Hwa-Seung;Bang, Sun-Hwi;Lee, Young-Min;Lee, Yeon-Weol;Son, Chang-Gue;Cho, Chong-Kwan
    • The Journal of Internal Korean Medicine
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    • v.28 no.2
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    • pp.304-320
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    • 2007
  • Objective : To evaluate the effect of Astragalus Membranaceus Extrac (AME) on myelosuppression, activity and immune modulation in 5-fluorouracil (5-FU) treated mice. Method : We carried out complete blood count, histological analysis of bone marrow, and cell colony forming assay for hematopoietic progenitor to evaluate the effect of AME on myelosuppression and conducted swimming test, survival rate, nitric oxide (NO) assay, 51Cr release assay in natural killer cell, mRNA expression of $IL-1{\beta}$, IL-2, IL-4, IL-6, IL-10, $TNF-{\alpht}$, $IFN-{\gamma}$, $TGF-{\beta}$ and GM-CSF in spleen cells to evaluate the effect of AME on quality of life (QOL). Results : AME improved 5-FU induced myelosuppression and peripheral blood count was recovered effectively, had significant efficacy to protect against chemotherapy induced marrow-destruction and on hematopoiesis compared with the control group, improved increase survival rate and the swimming time, had a stimulatory effect on macrophage activation and NK cell activity, and up-regulated cytokine gene transcription (IL-2, IL-6, $IFN-{\gamma}$) in murine immunologic system. Conclusion : We can conclude that AM is an effective herbal agent for improvement of myelosuppression and QOL in 5-FU treated mice.

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Gene Expression of Detoxification Enzymes in Tenebrio molitor after Fungicide Captan Exposure (살진균제인 캡탄 처리 후 갈색거저리의 해독효소 유전자 발현)

  • Jang, Ho am;Baek, Hyoung-Seon;Kim, Bo Bae;Kojour, Maryam Ali Mohammadie;Patnaik, Bharat Bhusan;Jo, Yong Hun;Han, Yeon Soo
    • Korean journal of applied entomology
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    • v.61 no.1
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    • pp.155-163
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    • 2022
  • The application of fungicides is indispensable to global food security, and their use has increased in recent times. Fungicides, directly or indirectly, have impacted insects, leading to genetic and molecular-level changes. Various detoxification mechanisms allow insects to eliminate reactive oxygen species (ROS) toxicity induced by agrochemicals including fungicides. In the present study, we analyzed the mRNA expression levels of detoxifying enzymes in Tenebrio molitor larvae following exposure to non-lethal doses (0.2, 2, and 20 ㎍/µL) of a fungicide captan. Transcripts of peroxidases (POXs), catalases (CATs), superoxide dismutases (SODs), and glutathione-s-transferases (GSTs) were screened from the T. molitor transcriptome database. RT-qPCR analysis showed that TmPOX5 mRNA increased significantly 24 h post-captan exposure. A similar increase was noticed for TmSOD4 mRNA 3 h post-captan exposure. Moreover, the expression of TmCAT2 mRNA increased significantly 24 h post-treatment with 2 ㎍/µL captan. TmGST1 and TmGST3 mRNA expression also increased noticeably after captan exposure. Taken together, these results suggest that TmPOX5 and TmSOD4 mRNA can be used as biomarkers or xenobiotics sensors for captan exposure in T. molitor, while other detoxifying enzymes showed differential expression.

Detection of Auxotrophic Mutants form Valsa ceratosperma, the Causal Fungus of Apple Canker (사과나무 부란병균(腐爛病菌) Valsa ceratosperma에서의 Auxotrophic Mutants의 검출(檢出))

  • Hong, Yeon Gyu;Uhm, Jae Youl
    • Current Research on Agriculture and Life Sciences
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    • v.5
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    • pp.119-126
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    • 1987
  • This study was conducted to elucidate the most appropriate method to obtain auxotrophic mutants from Valsa ceratosperma, the causal fungus of apple canker, which may be used as a gene marker in detecting the transfer of the factors of avirulent strains to virulent strains. Among the 3 kinds of synthetic media tested, each have two formula for minimal and complete, the medium which has been used in study of Endothia parasitica (E. P medium) was turned out to be most appropriate for the growth of V. ceratosperma. A medium for single colony formation from pycnidiospore of this fungus was developed by adding 0.5% L - sorbose to the E. P minimal medium. The period of incubation in dark for preventing the photoreactivation after U. V irradiation was estimated as about 60hrs at which most of the spores become binucleate. Largest number of putative auxotrophs were obtained at about 50second of irradiation to the spores smeared on the medium for single colony formation, at which the survival rate of spores was 5 to 6 percent. With these method developed in this experiment, 161 isolates of putative auxotrophs were detected among which the nutrient requirement for 10 isolates were determined. Five out of 10 mutants were still virulent to apple tree and all but one could not sporulate.

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