• Title/Summary/Keyword: DnaB

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Molecular Authentication of Schisandrae Fructus and Analysis of Phylogenetic Relationship based on nrDNA-ITS sequences (nrDNA-ITS 분자마커를 이용한 오미자(五味子) 종 감별 및 기원분석 -ITS 염기서열을 이용한 오미자(五味子) 감별-)

  • Moon, Byeong-Cheol;Ji, Yun-Ui;Seo, Hyeong-Seok;Lee, A-Young;Chun, Jin-Mi;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.25 no.4
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    • pp.47-54
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    • 2010
  • Objectives : The original plant species of Schisandrae Fructus (O-mi-ja) is prescribed as Schisandra chinensis $B_{AILL.}$, in Korea, but S. chinensis $B_{AILL.}$ and S. sphenanthera $R_{EHD.}$ et $W_{ILS.}$ in China. Moreover, fruit of several other species in genus Schisandra also have been used as the same herbal medicines. To develop a reliable method for correct identification of Schisandrae Fructus and to evaluate the phylogenetic relationship of S. chinensis and its related species, we analyzed internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA (nrDNA). Methods : Twenty-four plant samples of three Schisandra species and one Kadsura species, S. chinensis $B_{AILL.}$, S. spenanthera $R_{EHD.}$ et $W_{ILS.}$, S. nigra $M_{ax.}$ and Kadsura japonica $D_{UNAL}$ were collected from each different native habitate and farm in Korea and China. The nrDNA-ITS region of each samples were amplified using ITS1 and ITS4 primer and nucleotide sequences were determined after sub-cloning into the pGEM-Teasy vector. Authentic marker nucleotides were estimated by the analysis of ClastalW based on the entire nrDNA-ITS sequence. Results : In comparative analysis of the nrDNA-ITS sequences, we found specific nucleotide sequences including indels (insertions and deletions) and substitutions to distinguish C. chinensis, S. spenanthera, S. nigra, and K. japonica. These sequence differences at corresponding positions are avaliable nucleotide markers to determine the botanical origin of O-mi-ja. Moreover, we evaluated the phylogenetic relationship of four plant species by the analysis of nrDNA-ITS sequences. Conclusions : These marker nucleotides would be useful to identify the official herbal medicines by the providing of definitive information that can identify each plant species and distinguish it from unauthentic adulterants for O-mi-ja.

Identification of host plant species of Balanophora fungosa var. indica from Phnom Bokor National Park of Cambodia using DNA barcoding technique (캄보디아 프놈보콜국립공원의 Balanophora fungosa var. indica의 숙주식물에 대한 DNA barcoding 기법을 통한 동정)

  • Kim, Joo Hwan;Won, Hyosig
    • Korean Journal of Plant Taxonomy
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    • v.43 no.4
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    • pp.252-262
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    • 2013
  • During the floristic survey on Phnom Bokor National Park, Kampot, Cambodia, we encountered Balanophora fungosa var. indica, which is a tropical holoparasitic plant. To identify its host species, we collected host roots and trees nearby and tried to identify them using DNA barcoding approach. We applied plastid rbcL and matK gene regions as DNA barcode markers, and successfully amplified and sequenced the markers from 15 host roots and seven tree samples. Obtained host root sequences were identified as Primulaceae, Celastraceae, Myrtaceae, and Oleaceae, while trees nearby are Oleaceae, Myrtaceae, Sapindaceae, Rosaceae, Clusiaceae, Ericaceae, and Lauraceae. At genus level, host species are identified as Myrsine, Euonymus, Syzygium, and Olea, but failed in species discrimination. Myrsine (Primulaceae) and Olea (Oleaceae) are reported here as host species of B. fungosa var. indica for the first time. Further sampling and comparative work, and DNA barcoding will help recognize the biodiversity of the area and host species of Balanophora, together with their evolution.

Comparative analysis on genome-wide DNA methylation in longissimus dorsi muscle between Small Tailed Han and Dorper×Small Tailed Han crossbred sheep

  • Cao, Yang;Jin, Hai-Guo;Ma, Hui-Hai;Zhao, Zhi-Hui
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.11
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    • pp.1529-1539
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    • 2017
  • Objective: The objective of this study was to compare the DNA methylation profile in the longissimus dorsi muscle between Small Tailed Han and Dorper${\times}$Small Tailed Han crossbred sheep which were known to exhibit significant difference in meat-production. Methods: Six samples (three in each group) were subjected to the methylated DNA immunoprecipitation sequencing (MeDIP-seq) and subsequent bioinformatics analyses to detect differentially methylated regions (DMRs) between the two groups. Results: 23.08 Gb clean data from six samples were generated and 808 DMRs were identified in gene body or their neighboring up/downstream regions. Compared with Small Tailed Han sheep, we observed a tendency toward a global loss of DNA methylation in these DMRs in the crossbred group. Gene ontology enrichment analysis found several gene sets which were hypomethylated in gene-body region, including nucleoside binding, motor activity, phospholipid binding and cell junction. Numerous genes were found to be differentially methylated between the two groups with several genes significantly differentially methylated, including transforming growth factor beta 3 (TGFB3), acyl-CoA synthetase long chain family member 1 (ACSL1), ryanodine receptor 1 (RYR1), acyl-CoA oxidase 2 (ACOX2), peroxisome proliferator activated receptor-gamma2 (PPARG2), netrin 1 (NTN1), ras and rab interactor 2 (RIN2), microtubule associated protein RP/EB family member 1 (MAPRE1), ADAM metallopeptidase with thrombospondin type 1 motif 2 (ADAMTS2), myomesin 1 (MYOM1), zinc finger, DHHC type containing 13 (ZDHHC13), and SH3 and PX domains 2B (SH3PXD2B). The real-time quantitative polymerase chain reaction validation showed that the 12 genes are differentially expressed between the two groups. Conclusion: In the current study, a tendency to a global loss of DNA methylation in these DMRs in the crossbred group was found. Twelve genes, TGFB3, ACSL1, RYR1, ACOX2, PPARG2, NTN1, RIN2, MAPRE1, ADAMTS2, MYOM1, ZDHHC13, and SH3PXD2B, were found to be differentially methylated between the two groups by gene ontology enrichment analysis. There are differences in the expression of 12 genes, of which ACSL1, RIN2, and ADAMTS2 have a negative correlation with methylation levels and the data suggest that DNA methylation levels in DMRs of the 3 genes may have an influence on the expression. These results will serve as a valuable resource for DNA methylation investigations on screening candidate genes which might be related to meat production in sheep.

Detection of Hepatitis B Virus DNA in Liver Grafts Obtained from HBsAb and HBcAb Positive Organ Donors (HBsAb와 HBcAb가 양성인 장기 공여자의 간조직에서 Hepatitis B Virus DNA의 발현)

  • Jung, Chang-Woo;Jang, Joo-Young;Kim, Kyung-Mo;Lee, Sung-Gyu
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.10 no.2
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    • pp.166-172
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    • 2007
  • Purpose: It has recently been reported that de novo HBV infection following liver transplantation is caused by grafts from HBcAb positive donors, and this phenomenon has been observed in one third of the liver transplant patients in our center. Therefore, we investigated the presence of HBV virus DNA in liver tissues obtained from HBcAb positive donors to determine the mechanism by which de novo HBV infection occurs. Methods: This study was conducted on 6 patients that were HBsAg negative, HBsAb positive, and HBcAb positive who were donors for liver transplantation between November 1997 and November 1998 at Asan Medical Center. We isolated DNA from a portion of liver biopsy tissues that were obtained during the operation, and then identified the surface and core region of HBV DNA using nested PCR. In addition, four children who received liver grafts from these donors were monitored to determine if they became afflicted with non-HBV related diseases while receiving prophylaxis consisting of short-term HBIG treatment and long-term treatment with an antiviral agent. Results: The surface antigen region was identified in all 6 donors and the core antigen region was observed in 4 of the 6 donors. However, no episodes of de novo HBV infection with prophylaxis were observed. Conclusion: The results of this study support the results of previous studies, which indicated that HBV infection may be the main cause of de novo HBV infection in patients that receive HBsAb positive and HBcAb positive donor grafts.

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H-NS binding on dicA promoter DNA inhibits dicA gene expression (dicA promoter DNA에 붙는 H-NS 단백질에 의한 dicA 유전자의 발현 조절)

  • Yun, Sang Hoon;Lee, Yonho;Lim, Heon M.
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.191-198
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    • 2019
  • H-NS binds to promoter DNA and works as a general transcription silencer. DicA protein, by binding to the promoter DNA of dicA, activates dicA expression and at the same time inhibits expression of dicF and dicB, thus, exerting cell division control in Escherichia coli. H-NS complexed with a nucleoid protein Cnu was known to be involved in dicA expression. However, the exact nature of H-NS binding to dicA promoter DNA and the consequences of H-NS binding in expression of dicA is not clear. In this study, we explored the DNA binding activity of H-NS on the promoter DNA of dicA and found that H-NS binding occurs exclusively to the dicA promoter DNA. We never observed, however, H-NS binding at the vicinity of the dicA promoter. Temperature dependent oligomerization of H-NS was observed during DNA binding and the Cnu protein enhances the oligomerization process of H-NS binding. In vivo measurement of dicA expression in an hns deleted strain showed that dicA expression increased. These results demonstrated that H-NS binds specifically to dicA promoter DNA and functions as a transcription silencer.

발암물질의 조기검색법 개발 및 Chemoprevention에 관한 연구

  • 이병무;윤여표
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.193-193
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    • 1994
  • 발암물질의 조기검색법 개발 및 chemoprevention연구의 일환으로 발암물질과 DNA 및 단백질의 공유결합체인 발암물질-DNA 및 -단백질 adduct를 연구하였다. 발암물질(예, 밴조피렌)-단백질 adduct에 관한 연구에서는 시료(단백질)에 soluble protease를 이용하는 간편하고 손쉬운 ELISA(Enzyme Linked Immunosorbent Assay)분석법을 확립했다. 발암물질(예,벤조피렌,아플라톡신 B1) -DNA 및 -단백질 adduct를 이용한 발암성 조기검색법의 개발을 Ames test 및 염색체이상시험과 비교 연구한 결과 본 연구에서 새로이 개발한 DNA 및 Protein-adduct형성 시험법은 저농도에서 고농도에 이르기까지 뚜렷한 용량-반응 관계를 나타냈으며 Ames test 및 Chromosomal test에서 일어날 수 있는 false positive나 false negative의 결과를 나타낼 우려가 없었다. 벤조피렌-DNA adduct를 이용한 chemoprevention 연구에서는 항산화제로 알려진 비타민 E,C 및 $\beta$-carotene을 시험한 결과 용량의존적으로 벤조피렌-DNA adduct 형성을 억제하였다.

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KISS Korea Computer Congress 2005 (DNA 컴퓨팅 연산 과정의 통계 물리적 예측)

  • Kim J.S.;Kim J.C.;Noh Y.K.;Lee D.Y.;Zhang B.T.
    • Proceedings of the Korean Information Science Society Conference
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    • 2005.07b
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    • pp.253-255
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    • 2005
  • DNA 연산 과정의 열역학적 통계 물리학적 예측 모델을 기술한다. 온도를 천천히 내리는 시험관에서의 DNA string 들의 결합은 Metropolis 알고리즘과 진화 연산의 일종인 simulated annealing 알고리즘으로 설명될 수 있다 본 논문에서는 정리 증명 문제를 통해 위의 통계 물리학적 모델이 DNA 연산에 적용될 수 있음을 보인다. 여섯 종류의 DNA 가닥들의 시뮬레이션 결과와 온도에 대한 실험적인 fluorescence intensity의 비교를 통해 이 모델이 유효함을 보인다. 또한 목표 DNA 개수를 시뮬레이션으로 예측하고 그 결과를 electrophoresis gel image 와 비교하였다.

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DNA Barcoding of Boccardiella hamata (Annelida: Polychaeta: Spionidae) in South Korea

  • Lee, Geon Hyeok;Yoon, Seong Myeong;Min, Gi-Sik
    • Animal Systematics, Evolution and Diversity
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    • v.36 no.3
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    • pp.268-273
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    • 2020
  • A spionid polychaete, Boccardiella hamata (Webster, 1879) has been found from mud in crevices between the shells of oysters and adherent substrates in South Korea. The sequences of mitochondrial DNA (mtDNA) cytochrome c oxidase subunit 1 (CO1), 16S ribosomal DNA (16S), and the nuclear 18S ribosomal DNA (18S) from Korean individuals of Boccardiella hamata were determined in the present study. The molecular analysis based on the 18S rRNA gene sequences showed clear separation among the spionid polychaete species, and the sequences of Korean and Japanese individuals are completely identical. The morphological diagnosis and photographs of B. hamata are also provided.

The Protective and Inhibitory Effect of Antioxidants Found in Broussonetia kazinoki Siebold against Oxidative DNA Damage

  • Jang, Tae-Won;Choi, Ji-Soo;Kim, Hoi-Ki;Lee, Eun-Ja;Lee, Ki-Beom;Kwon, Tae-Hyung;Kim, Do-Wan;Ahn, Jeong-Jwa;Park, Jae-Ho
    • Korean Journal of Plant Resources
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    • v.32 no.6
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    • pp.714-722
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    • 2019
  • Oxidative DNA damage negatively affects humans and the research is currently ongoing to find ways to reduce oxidative stress. Oxidative stress has been identified as a key factor in triggering various diseases. Thus, its alleviation is important for human health. Broussonetia kazinoki (B. kazinoki) has been used in traditional Korean medicine as a dermatological therapy to treat burns, pruritus, and acne. B. kazinoki is generally segregated into peeled root (PR), root bark (RB), peeled stem (PS), and stem bark (SB). To assess these components for their antioxidant activity and protection against DNA damage, their ethyl acetate fractions were examined by 1,1-diphenyl-2-picryl hydrazyl (DPPH) and 2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radical scavenging assay. As a result of confirming the expression of factors involved in attenuating DNA damage, the protective effect of SB on oxidative stress suppressed the expression of p-p53 and γ-H2AX. Additionally, the levels of p53 and H2AX mRNA were significantly downregulated. In conclusion, these results indicated that the SB component of B. kazinoki had the potential to be used as an effective natural antioxidant compared to the other parts of the plant.

Cloning and Expression of cDNA Encoding a Cysteine Protease Inhibitor from Clamworm and Its Possible Use in Managing Anoplophora glabripennis Motschulsky (Coleoptera: Cerambycidae)

  • Li, Shengnan;Guo, Daosen;Zhao, Boguang;Ye, Jianling;Tian, Jie;Ren, Wenqing;Ju, Yunwei;Cui, Peng;Li, Ronggui
    • Journal of Microbiology and Biotechnology
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    • v.20 no.8
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    • pp.1243-1250
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    • 2010
  • A cDNA encoding a cysteine protease inhibitor (CPI) was isolated from the cDNA library of clamworm Perinereis aibuhitensis Grube. The deduced amino acid sequence analysis showed that the protein had 51%, 48%, and 48% identity with Zgc:153129 from Danio rerio, cystatin B from Theromyzon tessulatum, and the ChainA, stefin B tetramer from Homo sapiens, respectively. The gene was cloned into the intracellular expression vector pET-15b and expressed in Escherichia coli. The recombinant CPI (PA-CPI) was purified by affinity chromatography on Ni-charged resin and ion-exchange chromatography on DEAE-Sepharose FF. The relative molecular mass of PA-CPI was 16 kDa as deduced by SDS-PAGE. Activity analysis showed that the recombinant protein could inhibit the proteolytic activity of papain. A constitutive and secretive expression vector was also constructed, and the cDNA encoding CPI was subcloned into the vector for extracellular expression. Western blotting analysis results showed that the PA-CPI was secreted into the medium. Bioassay demonstrated that E. coli DH5${\alpha}$ harboring pUC18ompAcat-CPI showed a significant difference in mortality to the Asian longhorned beetle Anoplophora glabripennis compared with untransformed E. coli DH5${\alpha}$ and control.