• Title/Summary/Keyword: tail DNA

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Molecular Biological Studies on Korean Garlic Viruses

  • Choi, Jin-Nam;Song, Jong-Tae;Shin, Chan-Seok;La, Yong-Joon;Lee, Jong-Seob;Choi, Yang-Do
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 1994.06a
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    • pp.86-102
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    • 1994
  • To understand the molecular structure and pathogenesis mechanism of Korean garlic viruses, we have isolate cDNA clones for garlic viruses. The partial nucleotide sequences of 24 cDNA clones were determined and that of six clones containing poly (A) tail were compared with those of other plant viruses. One of those clones, V9 has 81.8% similarity in nucleotide sequence and 93.0% in deduced amino acid sequence, respectively, to the coat protein gene for garlic mosaic virus (GMV). Northern blot analysis with the clone V9 demonstrated that the genome of GMV is 7.8 kb long and has poly (A) tail. The anti-coat protein antibody for GMV recognizes 35 kDa polypeptide which could be the coat protein of GMV from infected garlic leaf extract or virus preparation. Clone G7 has about 62% of deduced amino acid sequence identity with the members of potyvirus group. Northern blot analysis with the clone G7 demonstrated that the genome of the potyvirus I garlic is 9.0 kb long and has poly (A) tail. The third clone, S81, shows 42% amino acid identity to the potexvirus. The other clones are under the characterization. To test the possibility of producing garlic virus resistant plant, we have designed a hairpin type ribozyme to cleave V9 RNA at the middle of the coat protein gene. From the cleavage reactions in vitro with two different sizes of RNA substrates, V9SUB (144 nucleotides) and V9 RNA (1,361 nucleotides), the ribozyme can cleave V9 sequence effectively at the predicted site. To study the activity of the ribozyme in vivo, plant transformation is in progress. Further possibilities to produce garlic virus resistant plant will be discussed.

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The Improvement of Chaga Mushroom (Inonotus Obliquus) Extract Supplementation on the Blood Glucose and Cellular DNA Damage in Streptozotocin-Induced Diabetic Rats (Streptozotocin으로 유발한 당뇨쥐에 있어서 차가버섯(Inonotus Obliquus)의 혈당 및 DNA 손상 개선효과)

  • Park, Yoo-Kyoung;Kim, Jung-Shin;Jeon, Eun-Jae;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.42 no.1
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    • pp.5-13
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    • 2009
  • Mushrooms have become a largely untapped source of powerful new pharmaceutical products that poses anti-inflammatory, and antimutagenic, and antioxidant activities. The antioxidant effects of the mushroom may be partly explained by protecting cellular components against free radical. The aim of this study was to investigate the protective effect of chaga mushroom against diabetes, via the mitigation of oxidative stress and reduction of blood glucose, in streptozotocin-induced diabetic rats. Rats were rendered diabetic by intravenous administration of STZ through tail at a dose of 50 mg/kg. Animals were allocated into four groups with 8 rats each. The control and diabetic control group were fed with standard rat feed. The other diabeic groups, the low chaga extract group and the high chaga extract group were fed ad libitum using 0.5 g/kg and 5 g/kg of chaga mushroom extract, respectively, for 4 weeks. The blood glucose levels in the two chaga extract groups showed a tendency to decrease but did not reach statistical significance after the supplementation. Leukocyte DNA damage, expressed as tail length, was found to be significantly lower in the high chaga extract group than in the diabetic control group (p > 0.05). Plasma level of total radical-trapping antioxidant potential (TRAP) was tend to be higher in the high chaga extract group compared with the diabetic control group. Erythrocyte antioxidant enzyme activities of two groups did not differ. Although we did not obtain beneficial effect on lowering blood glucose levels in the STZ-induced diabetic rats, this results suggest that the chaga mushroom extracts may initially act on protecting endogenous DNA damage in the short-term experiment.

Assessment of DNA Damage using an Alkaline Single Cell Gel Electrophoresis (SCGE) Comet Assay and Toxic Effects in Chickens by T-2 Toxin Treatment (T-2 toxin을 투여한 닭에서 Comet assay 방법을 이용한 DNA 손상 평가와 독성)

  • Hah Dae-Sik;Heo Jung-Ho;Lee Kuk-Cheon;Cho Myung-Heui;Kim Kuk-Hun;Kim Chung-Hui;Lue Jae-Du;Lee Seung-Hwan;Kim Gon-Sup;Kim Eui-Gyung;Kim Jong-Shu
    • Toxicological Research
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    • v.22 no.2
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    • pp.75-85
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    • 2006
  • This study was designed to evaluate the possible DNA damaging effects of T-2 toxin using an alkaline single cell gel electrophoresis (SCGE) comet assay and also to investigate toxic effects in chickens. A total of 20 chickens were used in these experiments. Graded concentrations of dietary T-2 toxin (0, 4, 8, and $16{\mu}g/g$ of diet) were given to groups of 5 broiler chickens. In comet assay, The DNA damage was analysed by the tail extent moment (TEM) and tail length (TL), which were used as markers of DNA strand breaks in SCGE. A significant dose-dependent increase in the extent of DNA migration as well as in the percentage of cells with tails was observed after treatment with T-2 toxin (P<0.05). Treatment with the low T-2 toxin ($4{\mu}/g$ of diet) induced a relatively low level of DNA damage in comparison with the high T-2 toxin ($16{\mu}/g$ of diet) group. The growth rate was significantly reduced by concentrations of 8, and $16{\mu}/g$ of diet (P < 0.05). The feed conversion ratio were significantly affected by any concentrations (P < 0.05). The relative weight of the spleen, and lung was decreased by the growth inhibitory concentrations. The bursa of Fabricius, thymus, and kid- ney were decreased in relative weight by concentrations of $16{\mu}/g$ of diet. The relative weight of the liver and heart were unaffected. The hemoglobin (Hb), hematocrit (HCT), and mean corpuscular hemoglobin (MCH) were decreased at concentration of $16{\mu}/g$ of diet. As compared with control chickens, there was no marked change in serum components except uric acid in T-2 treated chickens. All lymphoid tissues retained atrophic and lymphoid cell depletion throughout the three weeks trial.

Protective Effect of Kefir Grain Against Dextran Sodium Sulfate-Induced Colitis in Rats (Dextran Sodium Sulfate로 대장염을 유도한 흰쥐에서 캐피어 원말의 장보호 효과)

  • Ko, Young-Eun;Kim, Mi-Kyoung;Cho, Han-Young;Lee, In-Young;Ly, Sun-Yung
    • Journal of Nutrition and Health
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    • v.41 no.5
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    • pp.391-401
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    • 2008
  • Probiotics have emerged as a potential treatment modality for numerous gastrointestinal disorders, including IBD. However, few probiotics have undergone appropriate preclinical screening in vivo. Kefir is considered a probiotic, benefiting the host through its effects in the intestinal tract. Despite numerous studies examining the action of probiotics on the host organism, few have analyzed the effects on intestinal environment. We assessed the protective effect of kefir for three weeks before inducing colitis with 2% dextran sodium sulfate for five days. The DSS loads were similar in all DSS treatment group. The results of the experiment are as follows. Food intake and FER of experimental groups were not significantly different each other, but water consumption tended to be higher in all DSS treatment groups as compared with the normal control. And visual inspection of feces revealed mild diarrhea in rat given 2% DSS. The anti-inflammatory activity of kefir was determined by myeloperoxidase activity during the DSS treatment, and there was no significant difference in any group. The levels of thiobarbituric acid reactive substances (TBARS) as a colonic lipid peroxidation were significantly lower in the kefir intake groups than in rats treated with 2% DSS alone. The DNA % in tail and tail moment values as a DNA damage level of the blood lymphocytes in kefir intake groups tended to be lower than 2% DSS treatment alone, especially tail lengths were significantly diminished. According to the colonic histopathological assay, there were a severe inflammation of lamina propria and submucosa and mild edema in mucosa and sub mucosa in DSS alone treated group. We found a slight regenerative change in kefir treatment groups. In our experiments, this means that ulcerative colitis related to oxidative injury might be prevented by kefir as a probiotic. Further studies of the potential benefits of kefir as a probiotic in inflammatory condition are encouraged.

Candidate Genes with Ovulation by Differential Display PCR in Small Tail Han Sheep

  • Liu, Shufang;Li, Hongbin;Song, Xuemei;Wang, Aihua;Wei, Caihong;Du, Lixin
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.9
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    • pp.1229-1233
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    • 2006
  • To find the candidate genes concerned with ovulation rate of sheep, Differential Display Reverse Transcription Polymerase Chain Reaction was employed to find the differently expressed cDNA controlling ovulation in the Small Tail Han sheep of polyembryony and in Tan sheep of single birth. Twenty-four primer pairs of three anchored primers and eight arbitrary primers were assembled to amplify the specialized bands from these sheep. Positive cross tests were applied to optimize the ascertainable PCR conditions in which different special bands can be identified by silver strain in one PCR tube. After eliminating the false positive PCR products by Northern hybridization, 24 differential display bands were acquired from the ovary in the Small Tail Han sheep. These EST bands were sequenced and 18 different ESTs were found in which five ESTs had several copies and 13 ESTs had only one copy. Comparing these ESTs with homologous sequences by BLAST in the GenBank, there were six ESTs with known open reading frame (ORF) and function, three ESTs with known ORF and no function, and 9 ESTs without homologous sequence. These ESTs partly represent several genes such as NOS2, tensin, TCRA, CDKN1A, ESR1 and ACTB which express especially in Small Tail Han sheep.

Nucleotide Sequence of Leghemoglobin cDNA from Canavalia lineata

  • An, Chung-Sun
    • Journal of Plant Biology
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    • v.37 no.2
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    • pp.167-173
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    • 1994
  • Poly(A)+ RNA was selected from Canavalia lineata root nodule RNA through oligo(dT) cellulose column and used for construction of a cDNA library using λgt10-EcoRI arms. The size of the library was 7.2$\times$105 pfu/mL. A full length leghemoglobin (Lb) cDNA clone, pCILb1(687 bp) isolated with soybean Lb probe, contained one open reading frame (ORF) of 447 bp with 54 bp plus 186 bp at 5' and 3' untranslated region, respectively. A consensus sequence of plant translation start region (AAAATGGG) was found at 5' untranslated region, and two polyadenylation-related sequence (AATAAA, AATAAG) and a conserved motif between them (gACTTGTT) were found upstream of poly(A)+ tail consisted of 13 (A)s at 3' untranslated region. The ORF encoded a polypeptide consisted of 149 amino acids with a molecular weight of 16.2 kD. Deduced amino acid sequences showed high degree of homology values with those of other Lbs ranging from 66% (Casuarina glauca) to 85% (Glycine max).

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Killing Activity and Molecular Properties of Bacteriophage Sigma FA1 of Bacillus circulans (Bacteriophage Sigma FA1의 치사활성과 구조특성)

  • 김철호;김동수
    • Microbiology and Biotechnology Letters
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    • v.19 no.6
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    • pp.553-560
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    • 1991
  • In the previous paper (10). a new temperate phage, Sigma FA1 had been isolated from B. circulans. Sigma FA1 had an icosahedral head with a diameter of about 70 nm, and a tail about 15 nm long, and beared a circularly permuted, linear duplex DNA. Signla FA1 killed sensitive cells by a single-hit process. Phage DNA injected into the cell immediately after infection was degraded slowly. Our results indicate that the killing action of Sigma FA1 is different from the phenomenon of abortive infection and suggest that the killing might be caused by a proteinaceous component of Sigma FA1.

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Characterization of an Easter Lily Calmodulin cDNA Clone (백합실물에서 하나으 Calmodulin cDNA 클론 연구)

  • Kim, Seong-Ryong;An, Gyu-Heung
    • Journal of Plant Biology
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    • v.39 no.1
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    • pp.9-13
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    • 1996
  • A clone, LCM1, which encodes calmodulin (CaM) was isolated and characterized from monocot lily (Lilium longiflorum Thunb.) plants. The clone is 681 bps and contains the 447 bp coding region, 8 bp leader sequence, 210 bp 3'-untraslated region, and a poly(A) tail. The coding region of 149 amino acids encodes a protein of predicted Mr 17 kD. Comparison of the LCM1 amino acid sequence with other CaMs revealed that the protein is highly conserved among various living organisms. The expression level of calmodulin gene in lily was studied by RNA blot analysis. The LCM1 mRNA was present in all tissues tested. However, a higher level of calmodulin was observed in anther and floral bud. The level of calmodulin mRNA in anther was about 10 times higher than that in anther was about 10 times higher than that in vegetative tissues. The anther preferential expression of CaM in lily is currently investigated in dicot plants.

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Polyadenylation Is Dispensable for Encapsidation and Reverse Transcription of Hepatitis B viral Pregenomic RNA

  • Lee, Hye-Jin;Lee, Jehan;Shin, Myeong-Kyun;Ryu, Wang-Shick
    • Molecules and Cells
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    • v.25 no.4
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    • pp.545-552
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    • 2008
  • A hepadnaviruses replicates its DNA genome via reverse transcription of an RNA template (pregenomic RNA or pgRNA), which has a cap structure at the 5' end and a poly(A) tail at the 3' end. We have previously shown that the 5' cap is indispensable for encapsidation of the pgRNA. A speculative extension of the above finding is that the cap contributes to encapsidation via its interaction with the poly(A) tail, possibly involving eIF4E-eIF4G-PABP interaction. To test this hypothesis, poly(A)-less pgRNAs were generated via cleavage by a cis-acting hepatitis delta virus ribozyme sequence. We found that accumulation of the poly(A)-less pgRNA was markedly diminished, mostly likely due to its reduced stability. Importantly, however, the remaining poly(A)-less pgRNAs were nonetheless encapsidated and reverse transcribed normally when the reduced stability was taken account. Our finding clearly contradicts the notion that the poly(A) tail has any function in encapsidation and viral reverse transcription.

Morphological and molecular characteristics of Paralecithodendrium longiforme (Digenea: Lecithodendriidae) adults and cercariae from Chinese pipistrelle bats and viviparid snails in Thailand

  • Thitichai Arttra;Pheravut Wongsawad;Chalobol Wongsawad;Nattawadee Nantarat;Preeyaporn Butboonchoo;Jong-Yil Chai
    • Parasites, Hosts and Diseases
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    • v.62 no.1
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    • pp.85-97
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    • 2024
  • This study aimed to describe the morphological and molecular characteristics of Paralecithodendrium longiforme (Digenea: Lecithodendriidae) adults and cercariae isolated in Thailand. Adult flukes were isolated from the Chinese pipistrelle bat (Hypsugo sp.), and cercariae were detected in the viviparid snail (Filopaludina martensi martensi) from Chiang Mai province. The morphological characteristics were observed and described using conventional methods, and the molecular characteristics with internal transcribed spacer 2 (ITS2) and 28S rDNA gene sequences. The adult flukes were fusiform, 0.84-0.98 mm in length, and 0.37-0.49 mm in width, and were distinguishable from other species by the presence of longitudinal uterine coils. The cercariae were nonvirgulate xiphidiocercariae, with the oral sucker bigger than the acetabulum, the tail without fin fold, a body size of 117.5-138.3×48.3-52.2 ㎛, and a tail size of 100.7-103.7×15.0-18.9 ㎛. Molecular studies revealed that the adults and cercariae shared 99.3% (ITS2) and 99.6% (28S rDNA) homology with each other. They were phylogenetically close to P. longiforme with an identity of 94.5% for ITS2 and 98.7% for 28S rDNA. This study provides new information on the natural definitive host and first intermediate host of P. longiforme in Thailand. The discovery of its cercarial stage in Filopaludina snails highlights the importance of monitoring the associated second intermediate host and prevention and control of this potentially zoonotic trematode.