• Title/Summary/Keyword: DNA-DNA hybridization

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Isolation and characterization of a novel short-chain alcohol dehydrogenase gene from Panax ginseng

  • Kim, Yu-Jin;Shim, Ju-Sun;Lee, Jung-Hye;Jung, Dae-Young;Sun, Hwa;In, Jun-Gyo;Yang, Deok-Chun
    • BMB Reports
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    • v.42 no.10
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    • pp.673-678
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    • 2009
  • The cDNA of alcohol dehydrogenase (PgADH) was isolated and characterized from the leaf of Panax ginseng. The cDNA had an open reading frame of 801 bp and a deduced amino acid sequence of 266 residues. The calculated molecular mass of the mature protein is approximately 29 kDa with a predicated isoelectric point of 6.84. Homology analysis revealed that the deduced amino acid of PgADH shares a high degree of homology with the short-chain ADH proteins of other plants. Genomic DNA hybridization analysis indicated that PgADH represents a multi-gene family. The expression of PgADH under various environmental stresses was analyzed at different time points using real-time PCR. ABA, SA and especially JA (80-fold) significantly induced PgADH expression within 24 h of treatment. The positive responses of PgADH to abiotic stimuli suggest that ginseng ADH may protect against hormone-related environmental stresses.

Differentially Expressed Genes by Methylmercury in Neuroblastoma cell line using suppression subtractive hybridization (SSH) and cDNA Microarray

  • Kim, Youn-Jung;Chang, Suk-Tai;Yun, Hye-Jung;Ryu, Jae-Chun
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.05a
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    • pp.187-187
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    • 2003
  • Methylmercury (MeHg), one of the heavy metal compounds, can cause severe damage to the central nervous system in humans. Many reports have shown that MeHg is poisonous to human body through contaminated foods and has released into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established so far. In this study, two methods, cDNA Microarray and SSH, were performed to assess the expression profile against MeHg and to identify differentially expressed genes by MeHg in neuroblastoma cell line. TwinChip Human-8K (Digital Genomics) was used with total RNA from SH-SY5Y (human neuroblastoma cell line) treated with solvent (DMSO) and 6.25 uM (IC50) MeHg. And we performed forward and reverse SSH method on mRNA derived from SH-SY5Y treated with DMSO and MeHg (6.25 uM). Differentially expressed cDNA clones were sequenced and were screened by dot blot and ribonuclease protection assay to confirm that individual clones indeed represent differentially expressed genes. These sequences were identified by BLAST homology search to known genes or expressed sequence tags (ESTs). Analysis of these sequences may provide an insight into the biological effects of MeHg in the pathogenesis of neurodegenerative disease and a possibility to develop more efficient and exact monitoring system of heavy metals as environmental pollutants.

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Thermococcus onnurineus sp. nov., a Hyperthermophilic Archaeon Isolated from a Deep-Sea Hydrothermal Vent Area at the PACMANUS Field

  • Bae, Seung-Seob;Kim, Yun-Jae;Yang, Sung-Hyun;Lim, Jae-Kyu;Jeon, Jeong-Ho;Lee, Hyn-Sook;Kang, Sung-Gyun;Kim, Sang-Jin;Lee, Jung-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1826-1831
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    • 2006
  • A novel hyperthermophilic, anaerobic, heterotrophic archaeon, designated strain $NA1^T$, was isolated from a deep-sea hydrothermal vent area (depth, 1,650 m) within the Papua New Guinea-Australia-Canada-Manus (PACMANUS) field. Cells of this strain were motile by means of polar flagella, coccoid-shaped with a diameter of approximately $0.5-1.0{\mu}m$, and occurred as single cells. Optimal temperature, pH, and NaCl concentration for growth were $80^{\circ}C$, 8.5, and 3.5%, respectively. The new isolate was an obligate heterotroph that utilized yeast extract, beef extract, tryptone, peptone, casein, and starch as carbon and energy sources. Elemental sulfur was required for growth and was reduced to hydrogen sulfide. The G+C content of the genomic DNA was 52.0 mol%. Phylogenetic analysis of the 16S rRNA gene indicated that strain $NA1^T$ belongs to the genus Thermococcus, and the organism is most closely related to T. gorgonarius, T. peptonophilus, and T. celer; however, no significant homology was observed among species by DNA-DNA hybridization. Strain $NA1^T$ therefore represents a novel species for which the name Thermococcus onnurineus sp. novo is proposed. The type strain is $NA1^T$ (=KCTC 10859, =JCM 13517).

Acinetobacter antiviralis sp. nov., from Tobacco Plant Roots

  • Lee, Jung-Sook;Lee, Keun-Chul;Kim, Kwang-Kyu;Hwang, In-Cheon;Jang, Cheol;Kim, Nam-Gyu;Yeo, Woon-Hyung;Kim, Beom-Seok;Yu, Yong-Man;Ahn, Jong-Seog
    • Journal of Microbiology and Biotechnology
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    • v.19 no.3
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    • pp.250-256
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    • 2009
  • Acinetobacter strain $KNF2022^T$ was isolated from tobacco plant roots during the screening of antiviral substances having inhibitory effects on Tobacco mosaic virus (TMV) and examined by phenotypic, chemotaxonomic, and genetic characterization. It was a nonmotile, Gram-negative bacterium. This strain contained Q-9 as the main respiratory quinone. The major cellular fatty acids of the isolate were 16:0, 18:1 w9c, and 16:1 w7c/15 iso 2OH. The DNA base composition was 44 mol%. Phylogenetic analysis based on the 16S rRNA sequence revealed that the isolate formed an evolutionary lineage distinct from other Acinetobacter species. Based on the evaluation of morphologic, physiologic, and chemotaxonomic characteristics, DNA-DNA hybridization values, and 16S rRNA sequence comparison, we propose the new species Acinetobacter antiviralis sp. nov., the type strain of which is $KNF2022^T$ (=KCTC $0699BP^T$).

Origin-related study of genetic diversity and heteroplasmy of Mongolian sheep (Ovis aries) using mitochondrial DNA

  • Kim, Yi Seul;Tseveen, Khaliunaa;Batsukh, Badamsuren;Seong, Jiyeon;Kong, Hong Sik
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.2
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    • pp.198-206
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    • 2020
  • Food and agricultural production sector, especially livestock production is vital for Mongolia's economic and social development. Domestic sheep play key roles for Mongolians, providing food (meat, milk) and raw materials (wool, sheepskin), but genetic diversity, origin of sheep populations in Mongolia have not been well studied. Studies of population genetic diversity is important research field in conservation and restoration of animal breeds and genetic resources. Therefore, this study aimed to investigate genetic characteristics and estimate origin through the analysis of mitochondrial DNA control region D-loop and Cytochrome b of Mongolian indigenous sheep (Mongolian native, Orkhon and Altanbulag) and one Europe sheep (Suffolk). As a result of there were found, 220 SNPs (Single nucleotide polymorphism) in the D-loop region, 28 SNPs in the Cytochrome B region, furthermore, 77 Haplotypes. The nucleotide diversity was only found in D-loop region (n = 0.0184). Phylogenetic analysis showed that 3 (A, B, and C) of 5 haplogroups of sheep have been identified in our research. Haplogroup C was only found in Mongolian indigenous sheep. Haplogroup D and E were not observed. As a result of haplogroups, haplogroup A was dominant (n = 46 of 94 sheeps), followed by haplogroup B (n = 36) and haplogroup C (n = 12). Sequence analysis showed that T deletion, insertion and heteroplasmy in D-loop region occurred at a high rate in Mongolian indigenous sheep population (T insertion = 47, T deletion = 83). The heteroplasmy, which has never been found in Mongolian sheep, has been newly discovered in this study. As a result, the Mongolian sheep varieties, which mainly derived from Asia, were in hybridization with European sheep varieties.

Detection of Prevotella intermedia and Prevotella nigrescens using Pn17 and Pn34 DNA Probes

  • Park, Chan-Ho;Kim, Pan-Soon;Kim, Hwa-Sook;Min, Jeong-Bum;Hwang, Ho-Keel;Jang, Hyun-Sun;Cho, Ki-Woon;Baek, Dong-Heon;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.35 no.1
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    • pp.13-19
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    • 2010
  • The DNA probes Pn17 and Pn34 were evaluated for their ability to specifically detect clinical strains of P. intermedia and P. nigrescens from a Korean population by dot blot hybridization. These probes were sequenced by extension termination and their specificity was determined by Southern blot analysis. The results revealed that the Pn17 sequence (2,517 bp) partially encodes an RNA polymerase beta subunit (rpoB) and that Pn34 (1,918 bp) partially encodes both rpoB (1-169 nts) and the RNA polymerase beta subunit (rpoB'; 695-1918 nts). These probes hybridized with both HindIII- and PstI-digested genomic DNAs from the strains of P. intermedia and P. nigrescens used in this study. Interestingly, each of the hybrid bands generated from the HindIII-digested genomic DNAs of the two bacterial species could be used to distinguish between them via restriction fragment length polymorphism. These results thus indicate that Pn17 and Pn34 can simultaneously detect P. intermedia and P. nigrescens.

Genome Mapping of an Extreme Thermophile, Thermus caldophilus GK24

  • Park, Jong Hoon;Park, Byung Chul;Koch, Suk Hoon;Kim, Joong Soo;Koh, Jeong Heon;Yang, Moon Hee;Kim, Yong Sung;Kim, Cheorl Ho;Kim, Myoung Hee;Kwon, Suk Tae;Lee, Dae-Sil
    • Genomics & Informatics
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    • v.1 no.1
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    • pp.50-54
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    • 2003
  • Genome of an extreme thermophile, Thermus caldophilus GK24 has been analyzed to construct the genomic map. The genomic DNAs encapsulated in agarose gel were digested with SspI, EcoRI, SpeI, and HpaI restriction endonucleases, and then the resulting genomic DNA fragments were analyzed by pulsed-field gel electrophoresis. Its restriction map has been constructed by analyzing sizes of the restriction fragments obtained from both complete and partial digestions. The circular form of its genome was composed of about 1.98 Mbp and a megaplasmid. The genomic loci for the genes of xylose isomerase, thioredoxin, tRNA-16S rRNA, 23S rRNA, L5 ribosomal protein, ADP-glucose pyrophosphorylase, DNA-ligase, and Tca DNA polymerase were determined by both Southern hybridization and PCR.

CAMVS(V1.0) : CGH Analyzer and Map Viewer using S-Plus(V1.0)

  • Kim, Sang-Cheol;Park, Chan-Hee;Seo, Min-Young;Jeong, Ha-Jin;Kim, In-Young;Chung, Hyun-Cheol;Rha, Sun-Young
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2004.11a
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    • pp.131-137
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    • 2004
  • DNA 단계에서의 유전자의 증폭과 소실은 종양의 발생과 진행에 중요한 역할을 한다. 유전자의 변화를 관찰하기 위해서 Comparative Genomic Hybridization(CGH) 기술이 많이 이용되어져 왔다. 최근에는 이러한 CGH 기술을 응용하여 cDNA microarray 를 이용한 고밀도 CGH(Microarray-CGH) 기술이 보고 되고 있다. Microarray-CGH 에서 유전자별 변화 정도를 유전자의 log-비의 값의 변화 정도와 염색체 위치 정보를 이용하여 DNA 단계에서의 유전자의 변화 정도를 확인 할 수 있다. 또한 동일한 유전자의 칩을 사용하여 RNA단계에서의 발현 양상과 직접 비교할 수 있는 장점이 있다. 현재 microarray 분석법은 많이 개발되고 실용화 되고 있으나 Microarray-CGH 분석을 위한 프로그램들은 아직 초보 단계며, 생물학자들이 사용하기 힘들고, 프로그램에 분석 자료를 적용하기 어려운 경향이 있다. 위와 같은 단점을 보완하기 위해서 개발된 CAMVS(V1.0) 프로그램은 S-plus(2000)을 기반으로 개발하였고, 복잡한 분석보다는 모든 결과들을 이미지화 할 수 있으며 파일로 결과를 쉽게 확인할 수 있도록 디자인하였다. CAMVS(V1.0)는 전체 염색체를 각 실험별로 비교 분석하는 부분, 특정 염색체를 특정 실험별로 비교 분석하는 부분과 실험간의 차이를 통계적으로 비교 분석하는 3 가지 카테고리로 구성되어 있다. 쉬운 알고리즘과 사용의 편리함, 분석결과의 다양한 그래픽, 새로운 알고리즘 추가의 용이성 등이 CAMVS(V1.0)가 가지고 있는 장점이며, Microarray-CGH를 분석하는데 아주 유용한 분석 도구이다.

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Occurrence of Natural Hybrid between Oplegnathus fasciatus and Oplegnathus punctatus from the South Sea of Korea (한국 남해에서 출현한 돌돔 (Oplegnathus fasciatus)과 강담돔 (Oplegnathus punctatus) 사이의 자연교잡종)

  • Kwun, Hyuck-Joon;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.22 no.3
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    • pp.201-205
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    • 2010
  • One specimen of a natural hybrid of an Oplegnathus (Oplegnathus fasciatus $\times$ Oplegnathus punctatus) was found in Tongyeong, Korea in August 2008. We, herein, describe its morphological and genetic characteristics and compare them with those of O. fasciatus and O. punctatus. In morphology, the hybrid showed many distinctive black rounded blotches on body sides and four faint vertical bars, being in those features similar to O. punctatus. Although the counts and measurements of the hybrid mostly overlapped between O. fasciatus and O. punctatus, the Oplegnathus hybrid resembled O. punctatus in the ratio of pelvic-fin length in standard length: Oplegnathus hybrid (26.7%) was closer to O. punctatus (26.4%) than to O. fasciatus (17.2~23.6%). In genetics, as a result of analysis of 510 base pair sequences of mitochondrial DNA 16S rRNA, the hybrid was closer to O. fasciatus (d=0.000~0.010) than to O. punctatus (d=0.020). Our results suggest that the natural hybridization represented by the subject specimen occurred between an O. fasciatus female and an O. punctatus male.

The Amount of Telomeres and Telomerase Activity on Chicken Embryonic Cells During Developmental Stages (닭의 발생 단계별 세포내 Telomere의 양적 분포양상과 Telomerase 활성도 분석)

  • Cho, E.J.;Choi, C.H.;Sohn, S.H.
    • Journal of Animal Science and Technology
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    • v.47 no.2
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    • pp.187-194
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    • 2005
  • Telomeres locate at the end of chromosomes and consist of a tandem repeat sequence of $(TIAGGG)^{n}$ and associated proteins. Telomerase is a ribonucleoprotein which act as a template for the synthesis of telomeric DNA. Telomeres are essential for chromosome stability and are related with cell senescence, apoptosis and cancer. This study was carried out to analyze the amount of telomeres and telomerase activity of chicken cells during embryonic and developmental stages. The whole embryos and prenatal tissues such as brain, heart, liver, kidney and testis at different developmental stages were obtained from Korean Native Chicken. The amount of telomeres on embryonic cells was analyzed by quantitative fluorescence in situ hybridization (Q-FISH) techniques using the chicken telomeric DNA probe. Telomerase activity was measured by telomeric repeat amplification protocol (TRAP) assay. Results indicated that the amounts of telomeric DNA on the most embryonic cells were gradually decreased during ontogenesis. Furthermore, the quantity of telomeres was quite different among embryonic tissues according to developmental origin. The relative amount of telomeres has more in regenerative cells such as embryonic disc and testicular cells than in non-regenerative cells such as liver, brain, heart and kidney cells. Telomerase activity was also highly detectable in most chicken cells at early embryonic stages. After 9 days of incubation, however, the telomerase activitie W