• Title/Summary/Keyword: nucleic acid

Search Result 465, Processing Time 0.031 seconds

Use of RAPD-PCR(Random Amplified Polymorphic DNA-Polymerase Chain Reaction) Method for a Detection of Pathogenic Listeria monocytogenes (RAPD-PCR(Random Amplified Polymorphic DNA - Polymerase Chain Reaction) 방법을 이용한 Listeria monocytogenes의 검색)

  • Park Bum-Joon;Sihn Eon-Hwan
    • The Korean Journal of Food And Nutrition
    • /
    • v.17 no.3
    • /
    • pp.254-259
    • /
    • 2004
  • Rapid detection of foodbome pathogens is becoming increasingly important. The requirement for faster, more reliable tests has lead to the development of a wide range of rapid methods. Among these methods, the use of systems based on nucleic acid based detection has been increasing since they offer advantages of reduction in test time and more reliable detection or identification. Random Amplification Polymorphic DNA(RAPD) method has been used to fingerprint foodbome microorganisms; Listeria monocytogenes. In this study, 10-mer primer OPG-13(5'-CTCTCCGCCA-3') was used to generate RAPD-PCR for detection of pathogenic L. monocytogenes of Listeria spp. Among 20 primers tested, OPG-13 showed on acceptable result for the differentiation of a pathogenic Listeria from non-pathogenic microorganisms. Pathogenic Listeria, L. monocytogenes(ATCC 15313, 19111, 19112, 19113) showed two bands for 700 bp and 1,500 bp while non-pathogenic bacteria, L. ivanovii, L. grayi, L. murrayi, L. innocua, L. welshimeri, and L. seeligeri had only one band sizing from 2,000 to 2,300 bp. This RAPD method proved to be a valuable to gain important information on sources of pathogenic bacteria in food industry.

Proteome characterization reveals the role of pollen and pistil of W22 (ga1; Ga1) in maize

  • Roy, Swapan Kumar;Yu, Jin;Kamal, Abu Hena Mostafa;Kwon, Soo Jeong;Cho, Kun;Cho, Seong-Woo;So, Yoon-Sup;Woo, Sun Hee
    • Proceedings of the Korean Society of Crop Science Conference
    • /
    • 2017.06a
    • /
    • pp.129-129
    • /
    • 2017
  • The first key point to the successful pollination and fertilization in plants is the pollen pistil interaction, referring to the cellular and molecular levels, which mainly play active roles in limiting gene flow among maize populations and between maize and teosinte. This study was carried out to identify proteins and investigate the mechanism of gametophytic factors using protein analysis. W22 (ga1); which didn't carry a gametophytic factor and W22 (Ga1), a near iso-genic line were used for the proteome investigation. SDS-PAGE was executed to investigate proteins in the pollen and pistil of W22 (ga1) and W22 (Ga1). A total of 44 differentially expressed proteins were identified in the pollen and pistil on SDS-PAGE using LTQ-FTICR MS. Among the 44 proteins, a total of 24 proteins were identified in the pollen of W22 (ga1) and W22 (Ga1) whereas 20 differentially expressed proteins were identified from the pistil of W22 (Ga) and W22 (Ga1). However, in pollen, 2 proteins were identified only in the W22 (ga1) and 12 proteins only in the W22 (Ga1) whereas 10 proteins were confirmed from the both of W22 (ga1) and W22 (Ga1). In contrary, 10 proteins were appeared only in the pistil of W22 (ga1) and 7 proteins from W22 (Ga1) while 3 proteins confirmed in the both of W22 (ga1) and W22 (Ga1). Moreover, the identified proteins were generally involved in hydrolase activity, nucleic acid binding and nucleotide binding. These results help to reveal the mechanism of gametophytic factors and provide a valuable clue for the pollen and pistil research in maize. In addition, it might provide a comprehensive insight on the proteins that were involved in the regulation of pollen-pistil interaction.

  • PDF

Plant Immunity against Viruses: Moving from the Lab to the Field (식물바이러스 면역반응 최신 연구 동향 및 전망)

  • Kim, Nam-Yeon;Hong, Jin-Sung;Jeong, Rae-Dong
    • Research in Plant Disease
    • /
    • v.24 no.1
    • /
    • pp.9-25
    • /
    • 2018
  • Plant viruses cause significant yield losses and continuously threaten crop production, representing a serious threat to global food security. Studies on plant-virus interactions have contributed to increase our knowledge on plant immunity mechanism, providing new strategies for crop improvement. The prophylactic managements consist mainly following international legislations, eradication of infected plants, and application of pesticide to decrease the population of vectors. Hence, putting together the pieces of knowledge related to molecular plant immunity to viruses is critical for the control of virus disease in fields. Over the last several decades, the outstanding outcomes of extensive research have been achieved on comprehension of plant immunity to viruses. Although most dominant R genes have been used as natural resistance genes, recessive resistance genes have been deployed in several crops as another efficient strategy to control viruses. In addition, RNA interference also regulates plant immunity and contribute a very efficient antiviral system at the nucleic acid level. This review aims at describing virus disease on crops and summarizes current resistance mechanisms. Furthermore, we will discuss the current biotechnological approaches to control viral diseases and the future questions that are to be addressed to secure crop production against viruses.

Effects of Dietary Synthetic Amino Acid Supplementation in Korean Rockfish Fry Sebastes schlegeli (치어기 조피볼락에 있어서 사료내 합성아미노산 첨가 효과)

  • 김강웅;박건준;옥임호;배승철;최영준;신인수
    • Journal of Aquaculture
    • /
    • v.15 no.3
    • /
    • pp.157-163
    • /
    • 2002
  • Thirteen groups, each consisting of 25 juveniles (0.64 g) of the Korean rockfish, Sebastes schlegeli were reared in aquaria after period of one week conditioning. Each group was fed with one of the 13 diets at the rate of 7~10% body weight (on a dry-matter basis) 2 times a day for 6 weeks. Each diet was supplemented with one of the following amino acids at 3g/kg diet: Alanine (Ala), Arginine (Arg), Glycine(Gly), Glutamate (Glu), Histidine (His), Isoleusine(Ile), Lysine (Lys), Methionine (Met), Phenylalanine (Phe), Proline (Pro), Threonine (Thr), Tryptophan (Trp) or Valine (Val). Groups fed with Pro, Thr, Met or Gly-supplemented diet showed significantly (p<0.05) higher weight gain and faster specific growth rate than the groups fed on other diets, while those fed Pro, Thr, Met or Gly were not significantly different each other(P>0.05). Feed efficiency (FE) of fish fed Pro was significantly higher than those fed the other diets except that fed Thr (P<0.05). However, there was no significant difference between FE of fish fed Pro and Thr, and among FE of fish fed Thr, Met and Gly (P>0.05). The requirement of rockfish is higher for Met and Thr. Extra Pro and/or Gly may also stimulate biosynthesis of the nucleic acids (IMP, GMP) which are known as the feed stimulant in fish.

Manipulation of Rumen Fermentation by Yeast: The Effects of Dried Beer Yeast on the In vitro Degradability of Forages and Methane Production

  • Ando, S.;Khan, R.I.;Takahasi, J.;Gamo, Y.;Morikawa, R.;Nishiguchi, Y.;Hayasaka, K.
    • Asian-Australasian Journal of Animal Sciences
    • /
    • v.17 no.1
    • /
    • pp.68-72
    • /
    • 2004
  • The effects of the addition of yeast on in vitro roughage degradability and methane production were investigated in order to clarify the effects of yeast on the rumen microbes and to establish methods of rumen manipulation. Three roughages (whole crop corn, rice straw and Italian ryegrass) were incubated for 3, 6, 12 and 24 h with or without dried beer yeast following the method described by Tilley and Terry. Using the same method, these roughages were incubated with or without yeast extract, albumin or purified DNA. In vitro methane production was measured with or without dried beer yeast at 12 and 24 h. The degradability of yeast was found to be 57 and 80% at 12 and 24 h, respectively. The rate of degradation of fraction b was 6.16%/h. There was a significant increase in roughage degradability at 6 h (p<0.05), 12 h (p<0.05) and 24 h (p<0.01) by dried yeast addition. The degradability of all three roughages was higher in the samples treated with yeast extract than in the no addition samples except in the case of rice straw incubated for 12 h. Nevertheless, the magnitude of increment was smaller with the addition of yeast extract than without the addition of yeast. With the addition of purified DNA, there were significant increases in roughage degradability at 6 h (p<0.01), 12 h (p<0.01) and 24 h (p<0.05); however, higher degradability values were detected in the samples to which albumin was added, particularly at 6 h. If the degradability values of the no addition samples with those of samples containing yeast, yeast extract, DNA and albumin were compared, the largest difference was found in the samples to which yeast was added, although it is worth noting that higher values were observed in the yeast extract samples than in the DNA or albumin samples, with the exception of the case of rice straw incubated for 24 h. Methane production was significantly increased at both 12 and 24 h incubation. The increment of roughage degradation and methane production brought about by the addition of dried beer yeast to the samples was thought to be due to the activation of rumen microbes. Water soluble fraction of yeast also seemed to play a role in ruminal microbe activation. The increment of degradability is thought to be partially due to the addition of crude protein or nucleic acid but it is expected that other factors play a greater role. And those factors may responsible for the different effects of individual yeast on ruminal microbes.

Convenient Virion Capture (VC)/PCR for Tomato yellow leaf curl geminivirus Occurring on Tomato in Korea (우리나라 토마토에 발생한 토마토황화잎말림바이러스(Tomato yellow leaf curl geminivirus)의 초간편 Virion Capture(VC)/PCR 진단법)

  • Cho, Jeom-Deog;Kim, Tae-Seong;Kim, Ju-Hee;Choi, Gug-Seoun;Chung, Bong-Nam;Choi, Hong-Soo;Kim, Jeong-Soo
    • Research in Plant Disease
    • /
    • v.14 no.3
    • /
    • pp.233-237
    • /
    • 2008
  • Tomato yellow leaf curl virus (TYLCV), a newly reported Geminivirus from tomato, generated recently large economic losses in Korea. Development of a fast and precise genetic diagnosis technique for detecting TYLCV which Agricultural research and extension services can utilize easy and handy is very important to prevent yield losses. Virion Capture (VC)/PCR is a simple, accurate and economical genetic detection method without any works or commercial kits for the extraction of the nucleic acid from the infected plants. Primers of twenty two for detection of TYLCV were designed and tested with extracted total DNA or crude sap from tomato leaf infected with TYLCV and healthy plant. Nine primers for total DNA using conventional PCR and another 9 primers for VC/PCR were selected eventually. Primers of six having same specificity were selected from the two methods and tested with other Geminivirus, Tobacco leaf curl virus (TLCV) by VC/PCR. Finally specific primers of four were selected for detection of TYLCV using VC/PCR, and Deng (540, 541), a degenerate primer for Geminivirus reported in 1996, was also developed for VC/PCR.

Effects of Vincristine on the Epiphyseal Plate of the Rat Tibia (Vincristine이 흰쥐 경골의 골단연골판에 미치는 영향)

  • Chung, Woo-Min;Kim, Jong-Kwan;Kim, Won-Kyu;Chung, Ho-Sam
    • Applied Microscopy
    • /
    • v.27 no.3
    • /
    • pp.225-234
    • /
    • 1997
  • Vincristine, a kind of anticancerous drugs, interferes with development of microtubles and synthesis of nucleic acid and proteins in cells, and destructs cytoplasmic membrane so that mitosis of cancer cells is inhibited. Unfortunately these anticancerous effects by vioneristime are not limited to specific cancer cells, so several side effects are produced. This study was performed to explore the effects of vincristine on the fine structure of cytoplasmic organelles and cartilagenous matrix in proximal epiphyseal plate of the tibia in rat. The results were as follows: 1. Cisternae of rough endoplasmic reticulum (RER) were fragmented and sacculated, and membrane-bound ribocomes of RER were detached at 3 and 6 hours after vincristine treatment. Severely dilated, fagmented and sacculated cisternae of RER were found at 12 hours after vincristine treatment, and at 24 hours after vincristine treatment a few cisternae were framented and sacculated. At 72 hours after vincristine treatment cisternae of RER were parallely well arraged. 2. Golgi complex was atrophied at 3, 6, and 12 hours after vincristine treatment, while at 72 hours after vincristine treatment the cisternae of Golgi complex were made of 5-6 layers. 3. Mitochondria with disorganized mitochondrial cristae and outer membrane-losed mitochondria were found at 3 hours after vincristine treatment. At 6 and 12 hours after vincristine treatment mitochondria had possessed disorganized cristae, and a few mitochondria with disorganized cristae were. observed at 24 hours after vincristine treatment. While at 72 hours after vincristine treatment mitochondria were shown distinct cristae and double membranes. 4. Phagosome were begun to observe at 3 hourse after vincristine treatment, and at 24 hourse after vincristine treatment many phagosomes were found, while at 72 hours after vincristine treatment a few phagosomes were observed. 5. In the cartilagenous matrix large-sized matrix granules were decreased and collagen fibrils were dispersed at 3, 6, and 12 hours after vincristine treatment, while at 72 hours after vincristine treatment many large-sized matrix granules and numerous matrix it is suggested that although vincristine may induce the degenerative changes of the chondrocyte, resulting in changes of components of the cartilagenous matirx, these toxic effects may be regressed with time.

  • PDF

Effects of Danggwisayeokgaohsuyusaenggang-tang(當歸四逆加吳茱萸生薑湯) on Collagen-induced Arthritis in Mice (Collagen으로 유발된 생쥐의 관절염에 대한 당귀사역가오수유생강탕(當歸四逆加吳茱萸生薑湯)의 효과)

  • Kim, Min-Kyun;Oh, Min-Seok
    • Journal of Korean Medicine Rehabilitation
    • /
    • v.21 no.2
    • /
    • pp.63-85
    • /
    • 2011
  • Objectives : This study was carried out to know the effects of Danggwisayeokgaohsuyusaenggang-tang(hereinafter referred to DST) on arthritis induced by collagen on DBA/1 OlaHsd mice. Methods : For this purpose, DST was orally administered to mouse with arthritis induced by collagen II. Cytotoxicity, high performance liquid chromatograph(HPLC) analysis, arthritis index, value of immunocyte in draining lymph node and paw joint, cytokine were measured in vivo. Results : 1. The cytotoxicity against human fibroblast cells(hFCs) was not measured in any concentration. 2. In HPLC analysis, There are high peak patterns at 8 minute(min), 12 min, 35 min, 45 min. 3. The arthritis index was decreased significantly. 4. The degree of arthritis induced damage of joint of DST group is slight compared with control group in histopathologic observation(Hematoxylin and eosin stain(H&E), Masson's trichrome(M-T) staining). 5. In total cell counts of draining lymph node(DLN) and paw joint, the cells in DLN decreased significantly on DST 200 mg/kg and the cells in paw joint decreased significantly on 200 mg/kg and 50 mg/kg. 6. In DLN, $CD4^+/CD25^+$, $CD3^+/CD69^+$, major histocompatibility complex(MHC), class-II/$CD11c^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg $CD3^+/CD8^+$ cells decreased significantly on DST 200 200 mg/kg, $CD4^+$, $CD3^+/CD44^+$ cells decreased. 7. In paw joints, $CD4^+$, $CD11b^+/Gr-1^+$ cells decreased significantly on DST 200 mg/kg and 50 mg/kg. 8. In joints, levels of $IL-1{\beta}$, IL-6, $TNF-{\alpha}$, cyclo-oxygenase-2(COX-2), NOS-II were decreased on DST 200 mg/kg and DST 50 mg/kg. 9. In analysing of cytokine in CD3/CD28 activated spleen, IL-17 was decreased significantly, IL-4 was increased significantly $INF-{\gamma}$ was decreased on DST 200 mg/kg. 10. In analysing of cytokine in collagen activated spleen, IL-17 were decreased significantly, IL-4 was increased significantly. Conclusions : This results demonstrated that DST suppressed the inflammatory progression of collagen-induced arthritis(CIA) mice and supported further studies are required to survey continuously in looking for the effective substance and mechanism in the future.

FURTHER PURIFIED GINSENG EXTRACT FRACTION (D-O-ANA) FOR INSULIN RELEASE AND ITS MODE OF ACTION COMPARED WITH THE ISOLATED RESIDUAL COMPONENTS (인삼성분 D-O-ANa이 인슐린 분비에 미치는 영향 및 작용기전에 관한연구)

  • KIMURA Masayasu;SUZUKI Jun;WAKI Isami;KIMURA Ikuko;TANAKA Osamu;MATSU-URA Hiromichi
    • Proceedings of the Ginseng society Conference
    • /
    • 1984.09a
    • /
    • pp.191-197
    • /
    • 1984
  • A further purified fraction (D-O-ANa) was obtained from DPG 3-2 fraction of Ginseng Radix by complete removal of saponins, nucleosides, nucleic acid bases, amino acids, and sugars. D-O-ANa - induced insulin release was investigated to compare with that of DPG 3-2 and other isolated components. Among the sub fractions of DPG 3-2, D-O-ANa exhibited the most potent release of insulin with or without high concentrations of glucose, and it particularly enhanced the second phase of glucose-induced insulin release. DGP 3-2 potentiated significantly the glucose-induced insulin release from the isolated islets of diabetic mice at increasing concentrations of extracellular calcium ions (0.16 - 2.5 mM). A definite relationship was found between calcium $(^{45}Ca)$ uptake and insulin release. Ginsenoside $(G)-Rb_1\;and\;G-Rg_1$ did not enhance the glucose-induced insulin release. The effect of ginseng saponins was blocked by glucose (16.7 mM), being distinctly different from the glucose-potentiated effect of DPG 3-2. The insulin release effect of $G-Rg_1$ was unaffected by the presence or absence of extracellular $Ca^{2+}$ and theophylline. Adenosine also increased insulin release from isolated islets, but had no effect on perfused rat pancreas. Arginine stimulated insulin release less evidently than D-O-ANa, though arginineand adenosine-induced glucagon releases were more remarkable. In conclusion, D-O-ANa appears to be a major fraction in insulin release activity of ginseng and its mode of action may be related to $Ca^{2+}$ ion uptake. This physiological mechanism was distinct from that of the abnormal release induced by ginseng saponins.

  • PDF

Comparison of One-Tube Nested-PCR and PCR-Reverse Blot Hybridization Assays for Discrimination of Mycobacterium tuberculosis and Nontuberculous Mycobacterial Infection in FFPE tissues

  • Park, Sung-Bae;Park, Heechul;Bae, Jinyoung;Lee, Jiyoung;Kim, Ji-Hoi;Kang, Mi Ran;Lee, Dongsup;Park, Ji Young;Chang, Hee-Kyung;Kim, Sunghyun
    • Biomedical Science Letters
    • /
    • v.25 no.4
    • /
    • pp.426-430
    • /
    • 2019
  • Currently, molecular diagnostic assays based on nucleic acid amplification tests have been shown to effectively detect mycobacterial infections in various types of specimen, however, variable sensitivity was shown in FFPE samples according to the kind of commercial kit used. The present study therefore used automated PCR-reverse blot hybridization assay (REBA) system, REBA Myco-ID HybREAD 480®, for the rapid identification of Mycobacterium species in various types of human tissue and compared the conventional one-tube nested-PCR assay for detecting Mycobacterium tuberculosis (MTB). In conventional nested-PCR tests, 25 samples (48%) were MTB positive and 27 samples (52%) were negative. In contrast, when conducted PCR-REBA assay, 11 samples (21%) were MTB positive, 20 samples (39%) were NTM positive, 8 samples (15%) were MTB-NTM double positive, and 13 samples (25%) were negative. To determine the accuracy and reliability of the two molecular diagnostic tests, the one-tube nested-PCR and PCR-REBA assays, were compared with histopathological diagnosis in discordant samples. When conducted nested-PCR assay, 10 samples (59%) were MTB positive and seven samples (41%) were negative. In contrast, when conducted PCR-REBA test, three samples (17%) were MTB positive, 10 samples (59%) were NTM positive and four samples (24%) were negative. In conclusion, the automated PCR-REBA system proved useful to identify Mycobacterium species more rapidly and with higher sensitivity and specificity than the conventional molecular assay, one-tube nested-PCR; it might therefore be the most suitable tool for identifying Mycobacterium species in various types of human tissue for precise and accurate diagnosis of mycobacterial infection.