• Title/Summary/Keyword: Single strand

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Authentication of Salted-dried Fish Species Using Polymerase Chain Reaction-Single Strand Conformational Polymorphism and Restriction Analysis of Mitochondrial DNA

  • Kim, Joo-Shin;Chu, Kin Kan Astley;Kwan, Hoi Shan;Chung, Hau Yin
    • Fisheries and Aquatic Sciences
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    • v.11 no.3
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    • pp.133-139
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    • 2008
  • Molecular techniques, including restriction fragment length polymorphism(RFLP) and polymerase chain reaction-single strand conformational polymorph isms(PCR-SSCP), were developed to identify salted, dried threadfin(Eleutheronema tetradactylum) and white herring(Ilisha elongata) fish. Using PCR with universal primers, conserved 367-bp fragments of the cytochrome b gene were amplified from fresh fish samples and sequenced. The sequences were then searched for specific restriction sites. The digestion of the PCR products with the endonucleases AvaI, FokI, MboII, and MspI generated RFLP, which was used to identify the commercial products. Similarly, the amplified PCR-SSCP products were developed and the products tested. Overall, similar patterns were found in the majority of the fresh and processed products. Based on the results, both RFLP and PCR-SSCP were useful in determining and validating the authenticity of the fish species used to prepare the commercial salted, dried products. A similar approach can be applied to other species.

Analysis on Constituent Elements and Microstructure of Fiberglass Splint and Cast

  • Ham, Joo Hyun;Jung, Han Suk
    • Korean Journal of Materials Research
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    • v.31 no.8
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    • pp.433-438
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    • 2021
  • In this study, microstructural characteristics and constituent elements of fiberglass splint and cast are examined using a scanning electron microscope and an energy dispersive X-ray spectrometer. As observed by the scanning electron microscope, fiberglass splint and cast had a porous structure with many bundles of fiberglass textures well assembled. Spaces between bundles of the fiberglass splint are triangular or elliptical shaped and the long-axis diameter is measured at about 1 mm. The thickness of fiber bundles covered with plaster is measured at 600 ㎛ and the diameter of a single strand of fiberglass is up to 10 ㎛. The thickness of the fiberglass bundle of the fiberglass splint is measured at about 700 ㎛. Spaces between bundles are formed in the shape of triangles with gentle edges and long-axis diameter of up to 1.4 mm, which is larger than that of the splint. The thickness of a single strand of fiberglass of the plaster-coated cast is 11.5 ㎛, which is thicker than that of fiberglass of the splint. As a result of analyzing constituent elements of the fiberglass cast and the splint with an energy dispersive X-ray spectrometer, Ca, Si, and Al components are identically detected. This result shows that the fiberglass cast has a smoother surface with hardened plaster than the fiberglass splint. The thickness of the fiberglass bundle and the thickness of a single strand of the fiberglass are also larger than those of the fiberglass splint.

Primer RNA Synthesis by E. coli RNA Polymerase on the SSB-coated 229-nt ssi Signal of Lactococcal Plasmid pGKV21 (Lactococcal plasmid pGKV21의 SSB-coated 229-nt ssi signal 상에서 E. coli RNA polymerase에 의한 시발체 RNA 합성)

  • Jeong, Jin-Yong;Kim, Eun-Sil;Kim, Sam-Woong;Kang, Ho-Young;Bahk, Jeong-Dong
    • Journal of Life Science
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    • v.19 no.3
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    • pp.305-310
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    • 2009
  • Plasmid pGKV21 contains a 229-nucleotide (nt) single-strand DNA initiation (ssi) signal. Using asymmetric PCR, we prepared a small single-stranded (ss) DNA fragment of the ssi signal and, using the 229-nt ssDNA fragment, determined the requirements of RNA polymerase for priming and DNA-protein interaction. The ssi fragment prepared was able to generate primer RNAs with almost the same efficiency as the $M13{\Delta}lac182/229$ phage DNA. However, the cssi (complementary strand of the ssi signal) fragment could not synthesize primer RNAs. This result suggests that the 229-nt ssi signal functions in a strand specific manner. Gel retardation and DNase I footprinting demonstrated that the synthesized ssi fragment could interact with both E. coli RNA polymerase and SSB protein to synthesize primer RNA. In Escherichia coli [pWVAp], an addition of rifampicin resulted in an accumulation of ssDNA, indicating that the host-encoded RNA polymerase is involved in the conversion of ssDNA to double-stranded plasmid DNA.

A Study on the Axial Stiffness Prediction of Stand Using Analysis of Variance (분산분석을 이용한 스트랜드의 축강성 예측에 관한 연구)

  • Park, Yong-Dae;Yang, Won-Ho;Heo, Seong-Pil;Seong, Gi-Deuk
    • Transactions of the Korean Society of Mechanical Engineers A
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    • v.25 no.1
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    • pp.127-134
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    • 2001
  • Wire ropes are widely used in cable car, suspension bridge and elevator, etc. and composed of single or multi-layer strands. It is difficult to find out the characteristics of a strand or wire rope because of complicated geometry and contact condition. In this study, the axial stiffness is evaluated using finite element method and reliable finite element analysis model is presented, taking into consideration the convergence on the length. The axial stiffness predictive equation of a strand is developed using analysis of variance, which can be applicable for characterizing the relationship between load and displacement when the strand configuration is determined.

Radiation-induced DNA strand breaks in EL4 cells and mouse spleen lymphocytes (방사선에 의한 EL4 마우스 백혈병세포 및 정상 마우스 비장 임파구 DNA strand breaks의 측정)

  • Kim, Sung-ho;Kim, Tae-hwan;Chung, In-yong;Yoo, Seong-yul;Cho, Chul-koo;Chin, Soo-yil
    • Korean Journal of Veterinary Research
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    • v.31 no.3
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    • pp.329-335
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    • 1991
  • The filter elution technique was used to assay $^{60}Co$ $\gamma$ ray-induced DNA strand breaks(SB) in EL4 mouse leukemia cell and mouse spleen lymphocyte. The lymphocytes were stimulated with lipopolysaccharide (LPS, $20{\mu}g/ml$) to label $[^3H]$ thymidine. EL4 cells and lymphocytes in suspension were exposed at $0^{\circ}C$ to 0Gy, 1Gy, 5Gy, 10Gy or l5Gy for DNA single strand breaks(SSB) assay and 0Gy, 25Gy, 50Gy, 75Gy or 100Gy for DNA double strand breaks(DSB) assay of $^{60}Co$ radiation and elution procedure was performed at pH12.1 and 9.6. The number of DNA strand breaks increased with increasing doses of r rays. The strand scission factor(SSF) was estimated in each experiment (eluted volume 21ml). The slope of SSB EL4 cells was $0.01301{\pm}0.00096Gy^{-1}$ (n=5), the slope of SSB for lymphocytes was $0.01097{\pm}0.00091Gy^{-1}$ (n=5) and the slope of DSB for lymphocytes was $0.001707{\pm}0.0000573Gy^{-1}$ (n=5). Thus EL4 cells were more sensitive to induction of DNA SSB by ionizing radiation than lymphocytes (p<0.005). The ratio of slope of dose-response relationship (SSF versus dose) of lymphocytes DNA SSB as compared with the slope of DNA DSB was 6.4.

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Joint Interactions of SSB with RecA Protein on Single-Stranded DNA

  • Kim, Jong-Il
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.562-567
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    • 1999
  • Single-stranded DNA binding protein (SSB) is well-characterized as having a helix-destabilizing activity. The helix-destabilizing capability of SSB has been re-examined in this study. The results of restriction endonuclease protection assays and titration experiments suggest that the stimulatory effect of SSB on strand exchange acts by melting out the secondary structure which is inaccessible to RecA protein binding; however, SSB is excluded from regions of secondary structure present in native single-stranded DNA. Complexes of SSB and RecA protein are required for eliminating the secondary structure barriers under optimal conditions for strand exchange.

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Radiation Protective Effect of vitamin C and Cysteine on DNA Damage in Mice Splenic Lymphocytes by Single Cell Gel Electrophoresis Assay (단세포 겔 전기영동법을 이용한 생쥐 비장 림프구 DNA 손상에 대한 비타민 C 및 시스테인의 방사선 방어효과)

  • 천기정;김진규;김봉희
    • Environmental Analysis Health and Toxicology
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    • v.16 no.1
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    • pp.17-20
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    • 2001
  • The alkaline comet assay, employing a single-cell gel electrophoresis(SCGE), is a rapid, simple and sensitive technique for visualizing and measuring DNA damage leading to strand breakage in individual mammalian cells. The protecting effect of pretreatment with vitamin C and cysteine on the DNA damage of gamma ray was investigated in mice splenic lymphocytes. Vitamin C and cysteine were administered orally for five consecutive days before irradiation. Four week old ICR male mice were irradiated wish 3.5Gy of γ-radiation and were sacrificed 3 days later. Spleens were taken for DNA damage examination by Comet assay and the tail moments of DNA single -strand breaks in tole splenic lymphocytes were evaluated. The results show that pretreatment with vitamin C and cysteine were effective in protecting against DNA damage by gamma ray. Administration of antioxidants like vitamin C and cysteine to mice before irradiation was effective in reducing the tail moment of splenic lymphocytes DNA.

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DC Performance of $Nb_3$Sn Cable Joints with multi-interfaces (다수의 접합경계를 갖는 $Nb_3$Sn 케이블 접합부의 직류 저항 특성)

  • 이호진;김기백;연제욱;홍계원;김기만
    • Journal of the Korean Institute of Electrical and Electronic Material Engineers
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    • v.13 no.2
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    • pp.170-176
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    • 2000
  • The joints with multi-interfaces was expected to have low DC resistance compared with those with single interface. The small size joint specimens joined with Nb3Sn sub-cables were fabricated to investi-gate the DC performance in the range of 0 to 600A transport current without external magnetic field. The joints with multi-interfaces have a few n-Ohm resistance, which is much lower than that of single lap joint. Because the interfaces between sub-cables of multi-interfaced joint are more complicated than those of single-interfaced joint, the soldering condition between sub-cables is very effective on the joint DC resistance.

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