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Molecular Cloning and Characterization of a recA-like Gene Induced by DNA Damage from a Fluorescent Pseudomonas sp.

  • Ok Bong Kim;Na Young Kim;Jae Hoon Jeong;Si Wouk Kim;Hye Gwang Jeong;Seong Myeong Yoon;Jong Kun Park;Jung Sup Lee
    • Animal cells and systems
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    • v.3 no.2
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    • pp.229-236
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    • 1999
  • The recA gene plays a central role in genetic recombination and SOS DNA repair in Escherichia coli (E. coli). We have previously identified a 42 kDa RecA-like protein inducible by a variety of DNA damages from a fluorescent Pseudomonas strain sp. and characterized its inducible kinetics. In the present study, we cloned and characterized the gene encoding the RecA-like protein by immunological screening of Pseudomonas genomic expression library using polyclonal E. coli anti-RecA antibodies as a probe. From 10$^{5}$ plaques screened, five putative clones were finally isolated. Southern blot analysis indicated that four clones had the same DNA inserts and the recA-like gene was located within the 3.2 kb EcoRI fragment of Pseudomonas chromosomal DNA. In addition, the cloned recA-like gene was transcribed into an RNA transcript approximately 1.1 kb in size, as judged by Northern blot analysis. The cellular level of RNA transcript of the cloned recA-like gene was increased to an average of 5.15- fold upon treatment with DNA damaging agents such as ultraviolet (UV)- light, nalidixic acid (NA), methyl methanesulfonate (MMS), and mitomycin-C (MMC). These results suggest that the cloned gene is inducible by DNA damage similarly to the recA gene in E. coli. However, the cloned gene did not restore the DNA damage sensitivity of the E. coli recA-mutant.

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Image-Based Relighting Rendering System (영상 기반 실시간 재조명 렌더링 시스템)

  • Kim, Soon-Hyun;Lee, Joo-Haeng;Kyung, Min-Ho
    • Journal of the HCI Society of Korea
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    • v.2 no.1
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    • pp.25-31
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    • 2007
  • We develop an interactive relighting renderer allowing camera view changes based on a deep-frame buffer approach. The renderer first caches the rendering parameters for a given 3D scene in an auxiliary buffer with the same size of the output image. The rendering parameters independent from light changes are selected from the shading models used for shading pixels. Next, as the user interactively edits one light at one time, the relighting renderer instantly re-shades each pixel by updating the contribution of the changed light with the shading parameters cached in the deep-frame buffer. When the camera moves, the cache values should be re-computed because the currently cached values become obsolete. We present a novel method to synthesize them quickly from the cache images of the user specified cameras by using an image-based technique. This computations are all performed on GPU to achieve real-time performance.

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The Studies on Identification of Lophatherum gracile(淡竹葉) Using AFLP fingerprinting Methods (AFLP fingerprinting법을 이용한 담죽엽의 감별법 연구)

  • Shim, Young-Hoon;Seong, Rack-Seon;Park, Ju-Young;Kim, Ji-Yeon;Cho, Chang-Hee;Lee, Jong-Hwa;Hyeon, Seong-Ye;Kim, Sun-Ho;Kim, Dong-Sup;Jang, Seung-Yeup
    • Korean Journal of Pharmacognosy
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    • v.41 no.4
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    • pp.297-302
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    • 2010
  • Lophatheri Herba is the aerial part of Lophatherum gracile Bronghiart(淡竹葉, Gramineae). 25~75 cm in length. Stem: cylindrical with nodes, empty inside, externally pale yellowish green. Leaf: dehiscent of lanceolate lamina, shrunken and rolled, 5~20 cm long, 10~35 mm wide; surface: pale green ~ yellowish green, parallel-formed with veins of square reticulate, more distinct of appearance on the lower surface. Banbusae Caulis In Taeniam is the stringy strip derived from the stem with the peeled-off epidermis of Phllostachys nigra Munro var. henosis Stapf, and Phllostachys bambusoides Siebold et Zuccarini(竹葉, Gramineae). Irregular in size and shape, thin plane ~ strip-shaped, sometimes powdery, sometimes 1~3 mm thick. Outer surface: pale green ~ yellowish green, sometimes grayish white L. gracile and P. nigra have different origins although they show similar morphologic features. We were able to distinguish between L. gracile and P. nigra which are almost indistinguishable through this study. AFLP(Amplifide Fragment Length Polymorphism) was more suitable for identifying differences between L. gracile and P. nigra in comparison with other genetic analysis using chemical analysis. Therefore. molecular biological methods are believed to be useful for discovering origins of herbal medicines.

Mitochondrial DNA Variation in Oysters (Crassostrea gigas Thunberg and C. nippona Seki) Populations from Korea and Japan (한국 및 일본산 참굴 (Crassostrea gigas Thunberg)과 한국산 바위굴(C. nippona Seki) 의 미토콘드리아 DNA 변이)

  • 박미선;김상해
    • Animal Systematics, Evolution and Diversity
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    • v.11 no.2
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    • pp.235-242
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    • 1995
  • The nucleotide sequence variation of mitochondria1 DNA were investigated by eight restriction endonucleases from two oyster species, Crassostrea gigas collected from two localities of South Korea and one locality of Japan and C. nippona collected from one locality of South Korea. The total mtDNA size in the oyster, C. gigas, from the three localities was approximately 18 kb and that in C. nippona was 22 kb. The restriction fragment patterns of mtDNA in C. gigas from the three localities by BamHI, BgII, and XhoI digestions were identical to one another. The degree of mtDNA sequence divergence of C. gigas between the two localities in Korea was 2% and that between Korean and Japanese C. gigas was 5%. The amount of sequence divergence between the two species of oysters was 42%.

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Novel Preparation and Characterization of the α4-loop-α5 Membrane-perturbing Peptide from the Bacillus thuringiensis Cry4Ba δ-endotoxin

  • Leetachewa, Somphob;Katzenmeier, Gerd;Angsuthanasombat, Chanan
    • BMB Reports
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    • v.39 no.3
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    • pp.270-277
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    • 2006
  • Helices 4 and 5 of the Bacillus thuringiensis Cry4Ba $\delta$-endotoxin have been shown to be important determinants for mosquito-larvicidal activity, likely being involved in membrane-pore formation. In this study, the Cry4Ba mutant protein containing an additional engineered tryptic cleavage site was used to produce the $\alpha4$-$\alpha5$ hairpin peptide by an efficient alternative strategy. Upon solubilization of toxin inclusions expressed in Escherichia coli and subsequent digestion with trypsin, the 130-kDa mutant protoxin was processed to protease-resistant fragments of ca. 47, 10 and 7 kDa. The 7-kDa fragment was identified as the $\alpha4$-loop-$\alpha5$ hairpin via N-terminal sequencing and mass spectrometry, and was successfully purified by size-exclusion FPLC and reversed-phase HPLC. Using circular dichroism spectroscopy, the 7-kDa peptide was found to exist predominantly as an $\alpha$-helical structure. Membrane perturbation studies by using fluorimetric calcein-release assays revealed that the 7-kDa helical hairpin is highly active against unilamellar liposomes compared with the 65-kDa activated full-length toxin. These results directly support the role of the $\alpha4$-loop-$\alpha5$ hairpin in membrane perturbation and pore formation of the full-length Cry4Ba toxin.

Agrobacterium Mediated Transformation from Callus Pretreated with Particle Bombardment in Lilium lancifolium Thunb. (Particle Bombardment에 의해 전처리 된 참나리(Lilium lancifolium Thunb.) 캘러스의 Agrobacterium tumefaciens을 통한 형질전환)

  • Nam, Sang-Wook;Kim, Hei-Young
    • Journal of Plant Biotechnology
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    • v.31 no.1
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    • pp.13-17
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    • 2004
  • To improve transformation efficiency, the callus of Lilium lancifolium Thunb. were bombarded by particles coated with pIG 121 Hm which include NPT II and GUS genes, and then cocultivated with Agrobacterium tumefaciens EHA101 which contain pIG121Hm binary vector, carrying neomycin phosphotransferase (NPT II) and $\beta$-Glucuronidase (GUS) genes. Three days after cocultivation with Agrobacterium tumefaciens and particle bombardment, the callus clusters were transferred to MS medium containing 1mg/L 2,4-D, 0.1mg/L BAP, 100mg/L kanamycin and 200mg/L carbenicillin. Four weeks after transfer to the selection medium, GUS expression was detected and PCR analysis revealed the presence of NPT II fragment of the expected size (700 bp) in the transformed callus. The GUS expression from Agrobacterium-mediated transformants after particle bombardment increased to over 3-folds compared with that of callus cocultivated with Agrobacterium tumefaciens without particle bombardment. The callus clusters containing kanamycin resistant gene were transferred to MS medium containing 1mg/L NAA and 1mg/L BAP. Somatic embryos were developed in four weeks and microbulbs expressing GUS were formed.

Review of Acute Traumatic Closed Mallet Finger Injuries in Adults

  • Botero, Santiago Salazar;Diaz, Juan Jose Hidalgo;Benaida, Anissa;Collon, Sylvie;Facca, Sybille;Liverneaux, Philippe Andre
    • Archives of Plastic Surgery
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    • v.43 no.2
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    • pp.134-144
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    • 2016
  • In adults, mallet finger is a traumatic zone I lesion of the extensor tendon with either tendon rupture or bony avulsion at the base of the distal phalanx. High-energy mechanisms of injury generally occur in young men, whereas lower energy mechanisms are observed in elderly women. The mechanism of injury is an axial load applied to a straight digit tip, which is then followed by passive extreme distal interphalangeal joint (DIPJ) hyperextension or hyperflexion. Mallet finger is diagnosed clinically, but an X-ray should always be performed. Tubiana's classification takes into account the size of the bony articular fragment and DIPJ subluxation. We propose to stage subluxated fractures as stage III if the subluxation is reducible with a splint and as stage IV if not. Left untreated, mallet finger becomes chronic and leads to a swan-neck deformity and DIPJ osteoarthritis. The goal of treatment is to restore active DIPJ extension. The results of a six- to eight-week conservative course of treatment with a DIPJ splint in slight hyperextension for tendon lesions or straight for bony avulsions depends on patient compliance. Surgical treatments vary in terms of the approach, the reduction technique, and the means of fixation. The risks involved are stiffness, septic arthritis, and osteoarthritis. Given the lack of consensus regarding indications for treatment, we propose to treat all cases of mallet finger with a dorsal glued splint except for stage IV mallet finger, which we treat with extra-articular pinning.

Characterization of the Catabolite Control Protein (CcpA) Gene from Leuconostoc mesenteroides SY1

  • PARK JAE-YONG;PARK JIN-SIK;KIM JONG-HWAN;JEONG SEON-JU;CHUN JIYEON;LEE JONG-HOON;KIM JEONG HWAN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.749-755
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    • 2005
  • The ccpA gene encoding catabolite control protein A (CcpA) of Leuconostoc mesenteroides SYl, a strain isolated from kimchi, was cloned, sequenced, analyzed for transcript, and overexpressed in Escherichia coli. The ccpA ORF (open reading frame) is 1,011 bp in size, which can encode a protein of 336 amino acid residues with a molecular mass of 36,739 Da. The transcription start site was mapped at a position 49 nucleotides upstream of the start codon, and promoter sequences were also identified. The putative cre site overlapped with the -35 promoter sequence. The deduced amino acid sequence of the CcpA contained the helix-turn-helix motif found in many DNA-binding regulatory proteins. CcpA from 1. mesenteroides SY1 had $54.6\%$ identity with CcpA from Lactobacillus casei. The Northern blot experiment showed that ccpA was transcribed as a single 1.1 kb transcript, and transcription was repressed when grown on media containing glucose. CcpA was overproduced in E. coli BL21(DE3) cells using the pET expression vector, and purified to an apparent homogeneity. Gel Mobility Shift Assay with purified CcpA and a DNA fragment containing the ere sequence of the $\alpha$-galactosidase gene (aga) from L. mesenteroides SY1 revealed that CcpA bound specifically to the cre site of aga.

Expression of Human Serum Albumin in Milk of Transgenic Mice Using Goat β-casein/Human Serum Albumin Fusion Gene

  • Wu, H.T.;Chou, C.K.;Huang, M.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.6
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    • pp.743-749
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    • 2004
  • The gene encoding human serum albumin (HSA) was cloned from human liver cDNA library by PCR. The HSA cDNA in size of 2,176 bp, including 1,830 bp of open reading frame, was cloned into the plasmid carried with the 5'flanking sequence of goat $\beta$-casein gene (-4,044 to +2,025 bp) to get a tissue specific expression vector in mammary gland named pGB562/HSA (12.5 kb). A 9.6 kb DNA fragment in which the sequence is in order of goat $\beta$-casein gene regulatory sequence, HSA cDNA and SV40 polyadenylation signals was isolated from the pGB562/HSA by SacI and DraIII cutting, and used to microinject into the pronuclei of mouse fertilized eggs to produce transgenic mice. Three transgenic mice (2 female and 1 male) were identified by PCR and dot Southern blot analysis. The copy numbers of integrated transgene were more than 10 copies in line #21 and #26 as well as over 50 copies in line #31 of transgenic mice. HSA protein collected from the milk of lactating transgenic mice was confirmed by immuno-detection of Western and slot blot. The concentrations of HSA in the milk were from 0.05 to 0.4 mg/ml. An obvious antigen and antibody conjugate could be observed in immunohistochemical stain of mammary gland tissue from lactating day 11 of HSA transgenic mice. The transmission of transgene and its expression was recognized according to the results of RT-PCR and sequences analyses of their progeny.

Variations in Mitochondrial DNA of Pleurotus sajor-caju (여름느타리 버섯류의 미토콘드리아 DNA 비교)

  • Byun, Myung-Ok;Kim, Kyung-Soo;You, Chang-Hyun;Cha, Dong-Yeul
    • The Korean Journal of Mycology
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    • v.22 no.2
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    • pp.117-121
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    • 1994
  • Five strains of Pleurotus sajor-caju were collected from some countries including India and Papua New Guinea. Four strains produced brown fruitbody but the other strain from Papua New Guinea produced white fruitbody. Monokaryons obtained from both strains producing brown fruitbody and white fruitbody were mated each other. They showed different mating types such as brown strain of $A_1A_2B_1B_2$ and white strain of $A_3A_4B_3B_4$. DNAs were isolated from mycelia of five strains of P. sajor-caju. Mitochondrial DNA was seperated from nuclear DNA by bisbenzimide-CsCl ultracentrifugation. Digestion of the fungal mitochondrial DNA with Eco RI restriction endonuclease yielded from ten to fourteen fragments. Two strains of the five strains showed different restriction pattern of mitochondrial DNA from the other three strains. Summation of the fragment sizes gave a mitochondrial DNA size of about 60 to 65kb.

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