• 제목/요약/키워드: DNA isolation

검색결과 872건 처리시간 0.033초

Isolation of Dextran-producing Leuconostoc Zactis from Kimchi

  • Kim, Bong-Joon;Min, Bong-Hee;Kim, Jeongho;Han, Hong-Ui
    • Journal of Microbiology
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    • 제39권1호
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    • pp.11-16
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    • 2001
  • Tentative identification of Leuconostoc lactis IH23 isolated from kimchi (a fermented vegetable product) has been described previously with 16S rDNA sequencing (Choi, 1., M. Sc. Thesis Inha Univ.1999). This strain produced the slime identified as dextran based on IR, $\^$13/C- and $^1$H-NMR spectroscopic results. Further study proved that the isolate IH23 belongs to a homogeneous genetic group with L. lactis DSM 20202$\^$T/ and L. argentinum DSM 8581$\^$T/. The results showed DNA-DNA homology of 99-100%, 16S rDNA gene sequence similarity (97.7% ), and a phylogenetic relationship although L. argentinum DSM 8581$\^$T/ had lower homology (80-91%). These data indicate that L. argentinum DSM 8581$\^$T/ and the isolate IH23 belong to an identical species with L. lactis DSM 20202$\^$T/at the genetic level, although in carbohydrate fermentation, the isolate IH23 was mast closely related to L. argentinum DSM 8581$\^$T/ and quite different from L. lactis DSM 20202$\^$T/. Here we first report the isolation of consistent phenotypic variation in Leuconostoc lactis. We also emphasize that the nomenclature of subspecies needs to be differentiated into the three strains mentioned above in Leuconostoc lactis.

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Optimized Serological Isolation of Lung-Cancer-associated Antigens from a Yeast Surface-expressed cDNA Library

  • Kim, Min-Soo;Choi, Hye-Young;Choi, Yong-Soo;Kim, Jhin-Gook;Kim, Yong-Sung
    • Journal of Microbiology and Biotechnology
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    • 제17권6호
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    • pp.993-1001
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    • 2007
  • The technique of serological analysis of antigens by recombinant cDNA expression library (SEREX) uses autologous patient sera as a screening probe to isolate tumor-associated antigens for various tumor types. Isolation of tumor-associated antigens that are specifically reactive with patient sera, but not with normal sera, is important to avoid false-positive and autoimmunogenic antigens for the cancer immunotherapy. Here, we describe a selection methodology to isolate patient sera-specific antigens from a yeast surface-expressed cDNA library constructed from 15 patient lung tissues with non-small cell lung cancer (NSCLC). Several rounds of positive selection using patient sera alone as a screening probe isolated clones exhibiting comparable reactivity with both patient and normal sera. However, the combination of negative selection with allogeneic normal sera to remove antigens reactive with normal sera and subsequent positive selection with patient sera efficiently enriched patient sera-specific antigens. Using the selection methodology described here, we isolated 3 known and 5 unknown proteins, which have not been isolated previously, but and potentially associated with NSCLC.

Evaluation of DNA Extraction Methods from Low Copy Number (LCN) DNA Samples for Forensic DNA Typing

  • Eom, Yong-Bin
    • 대한의생명과학회지
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    • 제15권3호
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    • pp.229-232
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    • 2009
  • DNA isolation for PCR-based short tandem repeat (STR) analysis is essential to recover high yields of amplifiable DNA from low copy number (LCN) DNA samples. There are different methods developed for DNA extraction from the small bloodstain and gloves, commonly found at crime scenes. In order to obtain STR profiles from LCN DNA samples, DNA extraction protocols, namely the automated $iPrep^{TM}$ $ChargeSwitch^{(R)}$ method, the automated $QIAcube^{TM}$ method, the automated $Maxwell^{(R)}$ 16 DNA $IQ^{TM}$ Resin method, and the manual $QIAamp^{(R)}$ DNA Micro Kit method, were evaluated. Extracted DNA was quantified by the $Quantifiler^{TM}$ Human DNA Quantification Kit and DNA profiled by $AmpFISTR^{(R)}$ $Identifiler^{(R)}$ Kit. Results were compared based on the amount of DNA obtained and the completeness of the STR profiles produced. The automated $iPrep^{TM}$ $ChargeSwitch^{(R)}$ and $QIAcube^{TM}$ methoas produced reproducible DNA of sufficient quantity and quality trom the dried blood spot. This two automated methods showed a quantity and quality comparable to those of the forensic manual standard protocols normally used in our laboratory. In our hands, the automated DNA extraction method is another obvious choice when the forensic case sample available is bloodstain. The findings of this study indicate that the manual simple modified $QIAamp^{(R)}$ DNA Micro Kit method is best method to recover high yields of amplifiable DNA from the numerous potential sources of LCN DNA samples.

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홍조류인 한국산 김종에서의 염색체 DNA 분리방법 (Rapid and Efficient Purification of Nucleic Acids from the Macroalga Porphyra(Rhodophyta))

  • 류태형;최학선;최경희;이춘환
    • 한국식품영양과학회지
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    • 제27권6호
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    • pp.1166-1172
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    • 1998
  • A method for the isolation and purification of DNA from a red algae, Porphyra was innovated. The innovation of the method consists mainly of three steps that include sodium acetate treatment, chloroform extraction, and 0.2 volume isopropanol precipitation step. The sodium acetate treatment was designed to remove polysaccharide contamination, and the isopropanol step to remove proteins and salts contaminents. Genomic DNA,s of several species(for example, P. tenera, P. yezoensis, P. seriata, and P. pseudolinearis) was successfully isolated by the innovated method. The amount of DNA purified from one g of sample material with the innovated method was 53 g in average. The resulting DNA was characterized to include high molecular weight and showed no nuclease activity. The DNA was pure enough to be digested directly by various restriction enzymes without any difficulties. Porphyra DNA was pure enough and adequate for amplification reaction through the polymerase chain reaction (small nuclear rDNA PCR amplification).

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실리카로 코팅된 나노입자를 이용한 DNA 추출 방법 (DNA isolation using silica-coated magnetic nano particles)

  • 정승욱;이진성
    • 한국산학기술학회:학술대회논문집
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    • 한국산학기술학회 2007년도 춘계학술발표논문집
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    • pp.273-275
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    • 2007
  • 본 논문은 실리카 코팅 자성 나노입자를 이용한 새로운 DNA의 정제에 관한 것으로 기존의 negative-charged polyanion 형태의 수지 및 실리카 멤브레인 기술을 이용한 제품의 단점인 저속, 고비용의 정제방법을 개선할 수 있는 새로운 정제법을 기술한 것이다.

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상처 및 자외선 자극에 반응하는 여름느타리 cDNA 단편의 분리 및 그 발현 특성 (Isolation and Characterization of a Wound or UV Induced cDNA Fragment from Pleurotus sajor-caju)

  • 박수철;정욱진;정미정;김범기;유영복;류진창
    • 한국균학회지
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    • 제26권3호통권86호
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    • pp.314-320
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    • 1998
  • mRNA의 differential display방법에 의해 여름느타리 자실체의 상처 또는 자외선 처리시 발현되는 약 0.4kb의 cDNA 단편을 분리하였다. 이 cDNA 단편의 염기서열 분석결과 세포분열 촉진에 관여하는 cdc2-related protein kinase gene과 상당부분 유사성을 보였으며 RT-PCR 방법을 이용한 분리 유전자의 발현 실험을 통해 이 유전자가 정상 생장 환경에서는 갓, 대, 균사 등 모든 부위에서 기본적인 발현상태를 유지하고 있으며 기계적 상처 또는 자외선 처리에 의해 그 발현이 증폭됨을 확인하였다. 이러한 결과를 통해 향후 분리된 유전자의 연구를 통한 버섯 병 방어 관련 신호 전달 체계 분석에 대한 가능성을 검토해 보았다.

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Isolation and Characterization of cDNA Encoding Pyridoxal Kinase from Ovine Liver

  • Lee, Hyun-Shik;Choi, Soo-Young;Kwon, Oh-Shin
    • BMB Reports
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    • 제32권5호
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    • pp.502-505
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    • 1999
  • cDNA fragments of ovine liver pyridoxal kinase were amplified by PCR using degenerate oligonucleotide primers derived from partial amino acids sequences of the enzyme. Using PCR products as probes, several overlapping cDNA clones were isolated independently from an ovine liver and a human brain cDNA library. The largest cDNA clone for each was selected for sequence analysis. The ovine liver cDNA encodes a polypeptide of 297 amino acid residues with Mr of 32,925, whereas the human clone is comprised of an open reading frame encoding 312 amino acid residues with Mr of 35,102. The deduced sequence of the human brain enzyme is completely identical to that of human testes cDNA recently reported (Hanna et al., 1997). The ovine enzymes have approximately 77% sequence identity with the human enzyme although the two sequences are completely different in the N-terminus comprising 32 residues. This result suggests that pyridoxal kinase is highly homologous in mammalian species.

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