• 제목/요약/키워드: M13 DNA

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깽깽이풀의 핵형분석과 McFISH를 이용한 rDNA의 물리지도 작성 (Karyotype Analysis and Physical Mapping of rDNAs Using McFISH in Jeffersonia dubia Benth)

  • 김수영;최혜운;구달회;김찬수;방재욱
    • 한국약용작물학회지
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    • 제13권1호
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    • pp.48-51
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    • 2005
  • 보호식물이며, 약용식물인 깽깽이풀 (Jeffersonia dubia)을 대상으로 핵형 분석과 McFISH 기법을 이용한 염색체 분석을 수행하여 다음과 같은 결과를 얻었다. 체세포 염색체 수는 2n=2x=12였으며, 2쌍의 중부 염색체 (염색체 1, 3), 2쌍의 차중부 염색체 (염색체 2, 4) 그리고 2쌍의 차단부 염색체 (염색체 5, 6)로 구분되었고, 염색체의 평균 길이는 $1.95{\sim}3.50{\mu}M$이었다. McFISH기법을 이용하여 45S와 5S rDNA의 염색체상의 위치를 확인한 바, 3쌍의 45S rDNA signal은 4번, 5번 그리고 6번 염색체의 단완 말단 부위에서 관찰되었고, 한 쌍의 5S rDNA signal은 2번 염색체의 동원체 부위에서 관찰되었다.

Molecular Cloning and M13 Subcloning of Genes Encoding Catechol Dioxygenases

  • Kim, Young-Soo;Choi, Bong-Soo;Min, Kyung-Rak
    • Archives of Pharmacal Research
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    • 제15권1호
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    • pp.48-51
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    • 1992
  • Achromobacter xylosoxidans KF701 and Pseudomonas putida (NAH7) were significantly different in degradative capability of aromatic compounds including benzoates, biphenyls, and naphthalene. However, both of the bacterial strains can grown on catechol as the sole carbon and energy source. Catechol 2, 3-dioxygenase gene for naphthalene oxidation or biphenyl oxidation was cloned into Escherichia coli HB 701. A E. coli HB 101 clone containing catechol 2, 3-dioxygenase gene from P. putida (NAH7) contains a recombinant plasmid with 3.60kb pBR322 and 6-kb insert DNA. Another E. coli HB101 clone containing catechol 2, 3-dioxygenase gene from A. xylosoxidans KF 701 has a recombinant plasmid with 4.4kb pBR322 and 10-kb insert DNA. Physical maps of the recombinant plasmids were constructed, and catechol 2, 3-dioxygenase gene in the recombinant plasmide was further localized and subcloned int M13. The cloned-catechol 2, 3-dioxygenase game products were identified as yellow bands on nondenaturaing polyacrylamide gel after electrophoresis followed by activity staining with catechol solution.

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Cinnamic acid 유도체들의 SOS 반응을 지표로한 항돌연변이 효과에 관한 연구

  • 류재천;김승희;홍연탁;허문영
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1993년도 제2회 신약개발 연구발표회 초록집
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    • pp.174-174
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    • 1993
  • 목적: Cinnamic acid의 유도체들중에서 항돌연변이 효과가 보고된 바 있어, 본 연구에서는 E. coil PQ37균주를 사용한 SOS chromotest를 이용하여 약 170개 cinnamic acid의 유도체들의 항돌연변이 효과를 확인하고자 하였고, 1차로 확인된 수종의 유도체들에 대한 결과를 보고하고자 한다. 방법: SOS chromotest는 sul A:: lacz fusion strain 인 E. coli PQ37 에서의 chemical로 인한 DNA damage에 대한 SOS response를 $\beta$-galactosidase enzyme activity level로서 측정하는 실험이며, m-RNA 또는 protein synthesis에 대한 test chemicals의 cytotoxic저해효과를 알아보기 위하여 alkaline phosphatase를 병행측정하여 보완하였다. 결과 및 고찰: (-)S-9 경우는 4-nitroquinoline 1-oxide(4-NQO)를 유도물질로 하였으며 대략 Induction factor가 9.7 정도였고, (+)S-9의 경우는 aflatoxinB$_1$을 유도물질로 하였고 이때의 Induction factor는 13.8 정도였다. 이결과 1차로 test된 cinnamic acid유도체 6개중에서 RK001, RK002, RK003, RK004, RK005는 거의 항돌연변이 효과를 나타내지 않았으나, RK006은 항돌연변이 효과를 보여주어 이들의 Structure-Activity Relationship (SAR) 및 Dose-response와 RK006의 항돌연변이 효과의 mechanism에 관해 M13 mp2 viral DNA의 유전자 염기서열을 이용하여 항돌연변이 기전연구를 수행중이다.

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Voltammetric Assay of Mercury Ion in Fish Kidneys

  • Ly, Suw-Young
    • Toxicological Research
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    • 제24권1호
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    • pp.23-28
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    • 2008
  • Voltammetric analysis of mercury ions was developed using paste electrodes (PEs) with DNA and carbon nanotube mixed electrodes. The optimized analytical results of the cyclic voltammetry (CV) of the $1{\sim}14ng\;L^{-1}Hg(II)$ concentration and the square wave (SW) stripping voltammetry of the $1{\sim}12ng\;L^{-1}Hg(II)$ working range within an accumulation time of 400 seconds were obtained in 0.1 M $NH_4H_2PO_4$ electrolyte solutions of pH 4.0. For the relative standard deviations of the $1ng\;L^{-1}Hg(II)$, which were observed at 0.078% (n = 15) at the optimum conditions, the low detection limit (S/N) was pegged at $0.2ng\;L^{-1}(7.37{\times}10^{-13}M)$ for Hg(II). The results can be applied to assays in biological fish kidneys and wastewater samples.

First Report of Leaf Rust Caused by Puccinia caricis in Farfugium japonicum in Korea

  • Yun, Yeo Hong;Kwon, Hyuk Woo;Ahn, Hong Seok;Kim, Seong Hwan
    • Mycobiology
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    • 제43권3호
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    • pp.351-353
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    • 2015
  • Farfugium japonicum is used in traditional medicine and as an edible herb in China and Korea. In July 2013, leaf spots were observed in F. japonicum seedlings at Ulleung Island, Gyeongsangbuk Province, Korea. Early symptoms on the leaf adaxial surface included roughly circular yellow spots that later developed brown, necrotic centers. The aecia were hypophyllous, cupulate, yellowish, $180{\sim}430{\mu}m$ in diameter, clustered, and erumpent with a peridium with a recurved margin. The aeciospores were globoid, $14{\sim}17{\times}13{\sim}16{\mu}m$, light yellow or colorless, and densely verrucose. The 28S rDNA sequence of the isolate was identical to each other and shared 99% identity with Puccinia caricis. This is the first report of rust caused by P. caricis in F. japonicum in Korea or elsewhere in the world.

시중 유통 중인 한우와 수입쇠고기의 유전자 비교 및 위생 시험 (Genetic Comparison and Hygienical Test Between Korean Native Beef(Hanwoo) and Imported Beef(Holstein) Available in the Market)

  • 서정희;홍준배;정윤희;김말남
    • 한국식품위생안전성학회지
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    • 제13권4호
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    • pp.388-393
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    • 1998
  • 시장개방화로 수입쇠고기의 물량은 급증하여 최종 유통과정에서 수입쇠고기가 한우로 판매되어 소비자가 피해를 보는 등 사회적 물의를 일으키는 경우가 종종있다. 따라서 본 연구에서는 한우와 수입쇠고기의 과학적 판별을 위하여 시중에 유통 중인 한우와 수입쇠고기를 최근 유전자 기법인 PCR-RAPD를 이용하여 연구하였으며 아울러 위생에 대한 안전성 문제를 점검하기 위하여 장관출혈성 대장균과 식중독 세균에 대한 미생물 시험을 실시하였다. PCR 분석에 사용된 DNA 증폭조건은 $1{\times}Taq$ polymerase buffer, 1.5 mM $MgCl_2,\;50\;\mu\textrm{M}$ dNTP, 100ng primer, 2.5 unit Taq polymerase(perkin Elmer AmpliTaq), 5~20 ng template DNA이며 최종 반응용액은 $50\;\mu\textrm{\ell}$이다. 증폭산물의 크기는 대개 0.5 kbp~2.0kbp 사이의 범위에서 검출되었다. 수입 쇠고기에서만 DNA 크기가 약 1.2kbp인 유전자 표지인자가 확인되었으며 이 유전자 표지인자의 확인으로 한우와 수입쇠고기가 90%이상 구별되었으며 이는 한우 육간의 품종판별에 유용하리라 생각된다. 위생 시험 결과 최근 사회적으로 문제가 있었던 장관출혈성 대장균 O157:H7, O26, 등을 비롯하여 주요 식중독균인 Salmonella spp. 과 Listeria monocytogenes은 전혀 검출되지 않았다.

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Nucleotide Sequence of Leghemoglobin cDNA from Canavalia lineata

  • An, Chung-Sun
    • Journal of Plant Biology
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    • 제37권2호
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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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Introduction and Expression of Foreign Genes in Rice Cells by Particle Bombardment

  • Jeon, Jong-Seong;Jung, Hou-Sung;Sung, Soon-Kee;Lee, Jong-Seob;Choi, Yang-Do;Kim, Han-Jip;Lee, Kwang-Woong
    • Journal of Plant Biology
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    • 제37권1호
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    • pp.27-36
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    • 1994
  • For establishing a transformation system of rice, an efficient introduction of foreign genes into embryogenic cell suspension by particle bombardment was conducted. The particle inflow gun based on the acceleration of DNA-coated tungsten particles using pressurized helium was constructed for delivery of DNA into rice cells. Several bombardment parameters were optimized using the transient expression of GUS gene. The conditions that gave the highest GUS gene expression of about 1000 blue spots per g fresh weight of bombarded cells include treatment of the cells with 0.5 M osmotic pressure, and use of the 410 kPa helium, 110 mm target distance, 13 mm syringe filter holder and 5 $\mu$L DNA/tungsten mixtures. It was also confirmed that rice actin promoter-intron construct gave the highest expression of all promoter-sequences studied. Eight weeks after the bombardment, stably transformed calluses were obtained on the selection medium containing 100 mg/L G418 and showed the strong activity in in situ GUS assay.

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Construction of cDNA Library from Posterior Silk Gland (PSG) of Korean Oak Silkmoth, Antheraea yamamai and Molecular Cloning of Fibroin Heavy Chain Gene(FHC)

  • Lee, Jin-Sung;Kim, Soon-Jung;Kim, Ki-Hwan;Park, Young-Min;Suh, Dong-Sang
    • Journal of Life Science
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    • 제10권1호
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    • pp.10-13
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    • 2000
  • To develope the genetic source of oak wild silkworm, Antheraea yamamai, the cDNA library was constructed with poly A+ mRNA isolated from posterial silk gland of fifth instar larvae. Titer of the cDNA library was about 5.1$\times$105 pfu in total. We presumed that the titer covered almost all transcripts existed in Antherea yamamai. From cDNA library of Antheraea yamamai, fibroin heavy chain gene, which is specifically expressed from posterial silk gland of Antheraea yamamai, was screened using oligonuclotide probe specific to alanine rich motif of fibrin heavy chain gene of Antheraea pernyi. As a result, fibroin clones isolated from 5$\times$104 plaques showed the highest homolgy (95%) with that of Antherea pernyi in nucleotide of Anthereaea yamamai and Bombyx mori shows that there is no homologous sequence in the 3+ partial 채야후 region Genomic southern hybridization suggested that one copy is present. Northern hybridization showed that fibroin transcript was approximateely 9 kb in length.

DNA Fingerprinting of Red Jungle Fowl, Village Chicken and Broilers

  • Mohd-Azmi, M.L.;Ali, A.S.;Kheng, W.K.
    • Asian-Australasian Journal of Animal Sciences
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    • 제13권8호
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    • pp.1040-1043
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    • 2000
  • The genomic mapping of Red Jungle Fowl (Gallus gallus), local Village Chicken, and broiler was carried out by random amplified polymorphism DNA (RAPD) technique. Two different sets of arbitrary primers were used (Operon OPA01-20 and Genemed GM01-50). All the genomes of the three species of chickens were amplified with OPA01-20 primers. The genomes of the Red Jungle Fowl and local Village Chicken were further amplified with GM01-50 primers. Analysis of the results based on band sharing (BS) and the molecular size of individually amplified DNA fragments showed that Red Jungle Fowl and local Village Chicken shared the species similarity of 66% with Operon primers 01-20, 64% between local Village Chicken and broiler, and 63% when DNA bands between Red Jungle Fowl and broiler were compared. With GM01-50, the BS between Red Jungle Fowl and local village chicken increased to 72%. The results showed that the local village chicken is more closely related to Red Jungle Fowl than to broiler in the genetic distance. On the other hand, broiler is 1% closer in genetic distance to local village chicken than to Red Jungle Fowl. The results also indicated that primers like OPA-7, 8 and 9 can be used as species specific DNA markers for these three species of chickens.