• 제목/요약/키워드: ITS rDNA

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고추 역병 방제를 위한 저영양 길항세균 7F 균주의 동정 (Identification of the Oligotrophic Bacteria Strain 7F Biocontrolling Phytophthora Blight Disease of Red-pepper)

  • 김동관;여윤수;권순우;장길수;이창묵;이미혜;김수진;구본성;윤상홍
    • 식물병연구
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    • 제16권1호
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    • pp.41-47
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    • 2010
  • 고추 역병이 발생한 충주, 안면, 태안, 안동, 음성 그리고 괴산지역의 고추밭 토양에서 전체 10,753균주의 저영양 미생불을 분리하였다. 이들 중 Phytophthora capsid를 포함하여 식불체의 주요 병원성 곰팡이에 대해 길항작용을 보이는 균주들을 일차적으로 선발하였다. 최종적으로는 온실 고추유묘 검정을 통해 고추 역병에 대해 탁월한 방제가를 보이는 7F 균추가 선발되었다. 이 균주는 16SrDNA염기서열 분석과 API20 NE 키트를 통한 생리, 생화학적 분석을 통하여 Bacillus subtilis로 동정 되었다. 고추 역병균 포자를 처리한 고추 유묘 화분 검정에서, 처리25일 후 대조구는 고추 역병 이병 비율이 87%에 이르는 반면에 7F 균주를 처리한 처리구에서는 단지 6%의 이병율을 보였다.

Antioxidant and Apoptotic Effects of an Aqueous Extract of Urtica dioica on the MCF-7 Human Breast Cancer Cell Line

  • Fattahi, Sadegh;Ardekani, Ali Motevalizadeh;Zabihi, Ebrahim;Abedian, Zeinab;Mostafazadeh, Amrollah;Pourbagher, Roghayeh;Akhavan-Niaki, Haleh
    • Asian Pacific Journal of Cancer Prevention
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    • 제14권9호
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    • pp.5317-5323
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    • 2013
  • Breast cancer is the most prevalent cancer and one of the leading causes of death among women in the world. Plants and herbs may play an important role in complementary or alternative treatment. The aim of this study was to evaluate the antioxidant and anti-proliferative potential of Urtica dioica. The anti oxidant activity of an aqueous extract of Urtica dioica leaf was measured by MTT assay and the FRAP method while its anti-proliferative activity on the human breast cancer cell line (MCF-7) and fibroblasts isolated from foreskin tissue was evaluated using MTT assay. Mechanisms leading to apoptosis were also investigated at the molecular level by measuring the amount of anti and pro-apoptotic proteins and at the cellular level by studying DNA fragmentation and annexin V staining by flow cytometry. The aqueous extract of Urtica dioica showed antioxidant effects with a correlation coefficient of $r^2$=0.997. Dose-dependent and anti-proliferative effects of the extract were observed only on MCF-7 cells after 72 hrs with an $IC_{50}$ value of 2 mg/ml. This anti proliferative activity was associated with an increase of apoptosis as demonstrated by DNA fragmentation, the appearance of apoptotic cells in flow cytometry analysis and an increase of the amount of calpain 1, calpastatin, caspase 3, caspase 9, Bax and Bcl-2, all proteins involved in the apoptotic pathway. This is the first time such in vitro antiproliferative effect of aqueous extract of Urtica dioica leaf has been described for a breast cancer cell line. Our findings warrant further research on Urtica dioica as a potential chemotherapeutic agent for breast cancer.

김치 분리균인 Bacillus sp. JK-43이 생산하는 Cyclodextrin Glucanotransferase의 생산 및 특성 (Production and Characterization of Cyclodextrin Glucanotransferase fronm Bacillus sp. JK-43 Isolated from Kimchi)

  • 전홍기;배경미;김영희;백형석
    • 한국식품영양과학회지
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    • 제29권1호
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    • pp.41-48
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    • 2000
  • 김치 시료로부터 자동산화되지 않으며 열 및 중성 pH에서 안정한 AA 유도체인 AA-2G를 생산할 수 있는 당전이활성을 가진 CGTase 생산균주를 분리하였고, 분리균주의 형태학적, 배양학적, 생리학적 성질 및 16s-rDNA sequences를 조사한 결과 그람 양서의 간균으로 호기성이며 내생포자를 형성하는 전형적인 중온성 Bacillus sp. JK-43으로 동정되었다. Bacillus sp. JK-43의 CGTase는 AA-2G 뿐만 아니라 AA-6G로 추정되는 물질을 함께 생산하였으며, 효소 최적생산조건은 1.0% soluble starch, 1.0% yeast extract, 1.0% $Na_2CO_3\;0.1%\;K_2HPO_4,\;그리고\;0.02%\;MgSO_4{\cdot}7H_2O$가 함유된 배지에서 pH 7.0, $37^{\circ}C$에서 26시간 동안 진탕배양하였을 때였다. 각종 당공여채에 따른 Bacillus sp. JK-43의 AA-2G 생산성을 조사한 결과 ${\beta}-CD$에서 가장 높은 AA-2G 생산성을 보였으며, 식혜제도페액인 엿기름 및 밥당화액에서도 비교적 높은 AA-2G 생산성을 보였다. 또한 여러 가지 당수용체에 대한 JK-43의 CGTase의 당전이 반응을 검토한 결과 sucrose, mannitol 및 inositol에서 높은 당전이 수율인 70~90%를 나타내었다.

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벼(Orysa sativa L.) 뿌리로부터 분리된 내생 Streptomyces 균주의 동정 및 특성 (Identification and Characterization of an Endophytic Strain of Streptomyces from Rice Roots (Orysa sativa L.))

  • 김재헌;이준관
    • 미생물학회지
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    • 제47권4호
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    • pp.375-380
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    • 2011
  • 천안의 단국대학교 근처에서 채취된 벼 뿌리로부터 내생 방선균 한 주를 분리 동정하였다. 표면 살균된 벼 뿌리를 분리 배지 위에 올려 놓고 배양하면 4주 후에 뿌리 표면에 형성된 방선균 콜로니를 볼 수 있었다. 이 중에서 한 균주 JK-5를 분리하여 동정하였다. 16S rDNA 염기서열을 확인한 결과 이 분리균주 JK-5는 Streptomyces diastaticus subsp. ardesiacus와 동일한 균주였다. 형태학적 관찰과 화학분류학적 결과에 의해 분리균주 JK-5의 분류학적 위치를 확인하였다. 기중균사의 형태는 flexible하며 포자사슬은 타원형의 약 30개 정도의 매끄러운 포자로 이루어 졌다. 세포벽은 L,L-diaminopimelic acid를 포함하고 있었다. 당 분석에서는 특이적인 당이 발견되지 않았다. 지방산으로는 anteiso-15:0, anteiso-17:0, iso-16:0 가 주요 성분이었다. 퀴논 분석결과 MK-9 ($H_6$)와 MK-9 ($H_8$)이 주요 성분으로 나타났다. GC 함량은 72%로 측정되었다. 식물병원성 곰팡이에 대하여 비교적 높은 항진균활성을 보여 주었다. 벼에 감염된 분리균주 JK-5가 뿌리와 줄기에서 균사체를 형성하며 왕성하게 생장하는 것을 SEM을 통하여 확인하였다.

Accelerating Effect of $TNF-{\alpha}$ on the Rhus verniciflua-induced Growth Inhibition and Apoptosis in Human Osteosarcoma Cells

  • Kim, Hyun-Duck;Kook, Sung-Ho;Kim, Beom-Tae;Kim, Jong-Ghee;Jeon, Young-Mi;Lee, Jeong-Chae
    • Natural Product Sciences
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    • 제11권1호
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    • pp.45-49
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    • 2005
  • Previously, a flavonoid fraction, which consisted mainly of protocatechuic acid, fustin, fisetin, sulfuretin, and butein, here named RCMF [${\underline{R}}hus$ verniciflua Stokes (RVS) ${\underline{c}}hloroform-{\underline{m}}ethanol\;{\underline{f}}raction$], was prepared from a crude acetone extract of RVS which is traditionally used as a food additive and as an herbal medicine. In the present study, we investigated the effects of $TNF-{\alpha}$ on RCMF-induced growth inhibition and apoptosis induction using human osteosarcoma (HOS) cells. The results from tritium uptake and MTT assays showed that $TNF-{\alpha}$ treatment itself (10 ng/ml) did not induce any cytotoxicity, but it actively accelerated RCMF-mediated cytotoxicity of HOS cells. RCMF-induced cytotoxicity and its facilitation by $TNF-{\alpha}$ was verified to be apoptotic, based on the increased DNA fragmentation and low fluorescence intensity in nuclei after propidium iodide (PI) staining of HOS cells. This speculation was further demonstrated by monitoring the Annexin V/PI double staining which could discriminate the difference between apoptotic and necrotic deaths. Collectively, our findings indicate that $TNF-{\alpha}$ accelerates RCMF-induced cytotoxicity in HOS cells.

Growth hormone and receptor gene mutations in Chinese Banna miniature pig

  • Deng, J.Z.;Hao, L.L.;Li, M.T.;Lang, S.;Zeng, Y.Z.;Liu, S.C.;Zhang, Y.L.
    • Animal cells and systems
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    • 제15권4호
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    • pp.310-314
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    • 2011
  • The Banna miniature pig (BNMP) is a representative miniature pig breed in China. Even though BNMP dwarfism is obvious, its underlying causative mutations remain unknown. In this study, the BNMP and Large White pig (LWP) serum growth hormone (GH) and insulin-like growth factor (IGF-1) levels were detected by ELISA and compared. BNMP serum IGF-1 levels were significantly lower than LWP levels (P<0.05). The miniature condition may arise from mutations in the GH and GH receptor (GHR) genes. Therefore, GH and GHR cDNA from the BNMP were cloned into a pMD18-T vector by RT-PCR using the total RNA obtained from the BNMP's pituitary and liver tissues. Sequencing results indicated that the open reading frame of the BNMP GH gene is composed of a 26-residue signal peptide and a 191-residue mature peptide. The coding sequence of the BNMP GHR gene contained 639 amino acids, including a signal peptide that is 18 amino acids long. Two amino acid substitutions, A09V and R22Q, were found in the signal peptide of the GH gene. Additionally, the S104P mutation was found in the BNMP's mature GH protein. Four mutations in the cytoplasmic domain of GHR may influence the downstream signal transduction of GHR, which needs further experimental evidence.

Effects of alfalfa flavonoids extract on the microbial flora of dairy cow rumen

  • Zhan, Jinshun;Liu, Mingmei;Wu, Caixia;Su, Xiaoshuang;Zhan, Kang;Zhao, Guo qi
    • Asian-Australasian Journal of Animal Sciences
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    • 제30권9호
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    • pp.1261-1269
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    • 2017
  • Objective: The effect of flavonoids from alfalfa on the microbial flora was determined using molecular techniques of 16S ribosome deoxyribonucleic acid (rDNA) analysis. Methods: Four primiparous Holstein heifers fitted with ruminal cannulas were used in a $4{\times}4$ Latin square design and fed a total mixed ration to which alfalfa flavonoids extract (AFE) was added at the rates of 0 (A, control), 20 (B), 60 (C), or 100 (D) mg per kg of heifer BW. Results: The number of operational taxonomic units in heifers given higher levels of flavonoid extract (C and D) was higher than for the two other treatments. The Shannon, Ace, and Chao indices for treatment C were significantly higher than for the other treatments (p<0.05). The number of phyla and genera increased linearly with increasing dietary supplementation of AFE (p<0.05). The principal co-ordinates analysis plot showed substantial differences in the microbial flora for the four treatments. The microbial flora in treatment A was similar to that in B, C, and D were similar by the weighted analysis. The richness of Tenericutes at the phylum level tended to increase with increasing AFE (p = 0.10). The proportion of Euryarchaeota at the phylum level increased linearly, whereas the proportion of Fusobacteria decreased linearly with increasing AFE supplementation (p = 0.04). The percentage of Mogibacterium, Pyramidobacter, and Asteroleplasma at the genus level decreased linearly with increasing AFE (p<0.05). The abundance of Spirochaeta, Succinivibrio, and Suttonella at the genus level tended to decrease linearly with increasing AFE (0.05

Species identification and pathogenicity study of Colletotrichum isolates isolated from red-pepper and Chinese matrimony vine

  • Park, Eun-Sook;Lee, Bo-Heu;Min, Ji-Young;Cho, In-Joon;Kang, Beum-Kwan;Chung, Hae-Yeon;Yoo, Seung-Heon;Kim, Heung-Tae
    • 한국식물병리학회:학술대회논문집
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    • 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
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    • pp.131.1-131
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    • 2003
  • This study reports the identification of species of Colletotrichum strains originating from red-pepper and Chinese matrimony vine in Cheongyang. Ninteen isolates of red-pepper and 26 Coiletotrichum isolates of Chinese matrimony vine were compared with 5 isolates of strawberry representing C. gloeosporioides, by use of morphological and cultural criteria. Twenty three isolates among 26 isolates from Chinese matrimony vine were identified as C. acutatum, characterized by the low growth rates and the low sensitivity to carbendazim and diethofencarb. Also, all the isolates of red-pepper were identified as C. acutatum, showing the same characteristics as those of Chinese matrimony vine. Three and five isolates from Chinese matrimony vine and strawberry, respectively, were identified as C. gloeosporioides, characterized by the high growth rates and the high seneitivity to carbendazim and diethofencarb. There were differences in colony color and pathogenicity between Chinese matrimony vine isolates and red-pepper isolates of C. autatum. The isolates of C. acutatum from Chinese matrimony vine producing orange colored colonies with abundant spores showed the strong pathogenicity to Chinese matrimony vine, although they could not infect fruits of red-pepper by the wound inoculation. However, red-pepper isolates of C. acutatum producing gray colonies showed the strong pathogenicity to Chinese matrimony vine as well as red-pepper. Furthermore, comparative study on PCR amplification of ITS regions of rDNA was carried out using a number of Colletotrichum isolates. A species-specific primer could be used for the identification of C. acutatum from red-pepper and Chinese matrimony vine.

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Transcriptional Activator Elements for Curtovirus C1 Expression Reside in the 3' Coding Region of ORF C1

  • Hur, Jingyung;Buckley, Kenneth J.;Lee, Sukchan;Davis, Keith R.
    • Molecules and Cells
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    • 제23권1호
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    • pp.80-87
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    • 2007
  • Beet curly top virus (BCTV) and Beet severe curly top virus (BSCTV), members of curtoviruses, encode seven open reading frames (ORFs) within a ~3 kb genome. One of these viral ORFs, C1, is known to play an important role in the early stage of viral infection in plants during initiation of viral DNA replication. We used promoter:: reporter (${\beta}$-glucuronidase) gene fusions in transgenic Arabidopsis to identify the putative promoter region of BCTV ORF C1. Unlike other geminiviruses, the intergenic region of BCTV was not sufficient to promote C1 expression in transgenic plants. When sequences extending into the coding region of C1 were tested, strong expression of the reporter protein was observed in vascular tissues of transgenic plants. This expression was not dependent on the presence of the intergenic regions or proximal 5' portions of the C1 coding region. Transgenic plants expressing a reporter gene under control of the putative complete C1 promoter were inoculated with virus to determine if any viral transcript affected C1 expression. Virus inoculated plants did not show any altered pattern or change in of reporter gene expression level. These results suggest that (1) important transcriptional activator elements for C1 expression reside in the 3' portion of C1 coding area itself, (2) C1 protein does not auto-regulate its own expression and (3) C1 expression of two curtoviruses is controlled differently compared to other geminiviruses.

Physiological Characteristics and Production of Folic Acid of Lactobacillus plantarum JA71 Isolated from Jeotgal, a Traditional Korean Fermented Seafood

  • Park, Sun-Young;Do, Jeong-Ryong;Kim, Young-Jin;Kim, Kee-Sung;Lim, Sang-Dong
    • 한국축산식품학회지
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    • 제34권1호
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    • pp.106-114
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    • 2014
  • Folic acid, one of the B group of vitamins, is an essential substance for maintaining the functions of the nervous system, and is also known to decrease the level of homocysteine in plasma. Homocysteine influences the lowering of the cognitive function in humans, and especially in elderly people. In order to determine the strains with a strong capacity to produce folic acid, 190 bacteria were isolated from various kinds of jeotgal and chungkuk-jang. In our test experiment, JA71 was found to contain $9.03{\mu}g/mL$ of folic acid after 24 h of incubation in an MRS broth. This showed that JA71 has the highest folic acid production ability compared to the other lactic acid bacteria that were isolated. JA71 was identified as Lactobacillus plantarum by the result of API carbohydrate fermentation pattern and 16s rDNA sequence. JA71 was investigated for its physiological characteristics. The optimum growth temperature of JA71 was $37^{\circ}C$, and the cultures took 12 h to reach pH 4.4. JA71 proved more sensitive to bacitracin when compared with fifteen different antibiotics, and showed most resistance to neomycin and vancomycin. Moreover, it was comparatively tolerant of bile juice and acid, and displayed resistance to Escherichia coli, Salmonella Typhimurium, and Staphylococcus aureus with restraint rates of 60.4%, 96.7%, and 76.2%, respectively. These results demonstrate that JA71 could be an excellent strain for application to functional products.