• 제목/요약/키워드: Fragment size

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Direct Extraction of DNA from Soil for Amplification of 16S rRNA Gene Sequences by Polymerase Chain Reaction

  • Cho, Jae-Chang;Lee, Dong-Hun;Cheol, Cho-Young;Cho, Jang-Cheon;Kim, Sang-Jong
    • Journal of Microbiology
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    • 제34권3호
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    • pp.229-235
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    • 1996
  • Microgram quantities of DNA per gram soil were recovered with SDS- based and freeze-and thaw procedures. The average DNA fragment size was > 23 Kb. This method generated minimal shearing of extracted DNA. However, the DNA extracts still contained considerable amounts of humic impurities sufficient to inhibit PCR. Several approaches were used to reduce the interferences with the PCR (use of CTAF in extraction step, Elutip-d column purification, addition of BSA to PCR buffer) to accomplish PCR with DNA extract as a template. Most of the DNA extracts were not digested completely by restriction endonuclease, and CTAB-TREATED ane Elutip-d column purified DNA extracts were partially digested. Regarding as restriction enzyme digestion, all PCRs failed to amplify 16S rRNA gene fragments in the DNA extracts. In the case of DNA extracts only where BSA was added to PCR buffer, PCR was successfully conducted whether the DNA extracts were treated with CTAB or purified with columns. However, these two treatments were indispensable for humic impurity-rich DNA extracts to generate the PCR-compatible DNA samples. Direct extraction of DNA, coupled with these procedures to remove and relieve interferences by humic impurities and followed by the PCR, can be rapid and simple method for molecular microbiological study on soil microorganisms.

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Detection of Glycoproteins (B and D) and Thymidine Kinase Genes of Herpes simplex virus Type 2 Strain G

  • Kang, Hyun;Park, Jong-Kuk;Uh, Hong-Sun;Kim, Soo-Young;Lee, Hyung-Hoan
    • 대한바이러스학회지
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    • 제29권2호
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    • pp.99-105
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    • 1999
  • BamHI restriction pattern and genomic library of Herpes simplex virus type 2 (HSV-2) strain G were constructed, and locations of the glycoproteins gB and gD, and tk genes on the fragments were detected by Southern blot analysis. HSV-2 genomic DNAs were cleaved into twenty-seven fragments by BamHI enzyme in the range of 0.72 to 15.08 (total 150.44 kb), which were cloned into the BamHI site of pBluescript SK(+) to construct genome library of the HSV-2. The library was named by the order of the fragment size from smallest one to largest one. HSV-2 glycoprotein gD gene was located in pHLA2-21 and pHLA2-22 recombinant plasmids, gB gene in pHLA2-24 plasmid, and tk gene in pHLA2-11 clone by Southern blot analysis.

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Clostridium perfringens 감염에 의한 한우 집단 폐사 증례 (Mass mortality of Clostridium perfringens infection in Hanwoo)

  • 이정원;김지영;곽길한;육현수;손구례;고원석
    • 한국동물위생학회지
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    • 제34권1호
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    • pp.31-35
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    • 2011
  • This is a case report that 24 heads of cattle suddenly died without clinical signs in a Hanwoo farm. The cause of death was Clostridium perfringens enterotoxemia resulted from them with feeding leftover food. The clinical signs were observed just before the death; increase of heart rate, shallow and rapid respiration, amyostasia, spasm and so forth. In autopsy, blood coagulation disorder, a little abdominal inflation, hepatomegaly and different size of red spots, congestion hemorrhage and undercurrent of bloody exudation were observed in the entire parts of small and large intestines. C. perfringens were isolated from the substantive organs, and a unique fragment of 405bp C. perfringens was amplified by PCR. Therefore, this case was diagnosed as enterotoxemia caused by ${\alpha}$-toxin of C. perfringens A type.

Bacteriological detection of Brucella abortus and its characterization by PCR in the sporadic outbreak of bovine brucellosis in Gyeonggi province

  • Yang, Su-Jeong;Shim, Hang-Sub;Woo, Jong-Tae;Kim, Hye-Sung;Lee, Sung-Sik
    • 한국동물위생학회지
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    • 제30권2호
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    • pp.251-258
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    • 2007
  • Bovine brucellosis has occurred for years in Gyeonggi province under the national test and slaughter scheme. The serum agglutination test (SAT) is a diagnostic tool to confirm the disease despite the argument on its specificity. We selected 8 farms where only one or two individuals were diagnosed as brucellosis through SAT at the primary regular herd check and isolated the causative organism and characterized the species by species-specific PCR. The pathogen isolation was successful in 6 farms out of 8 farms by microbiological culture, showing the successful rate of 75%. The isolation rate of the causative organism represents 70% from supra-mammary lymph node and 60% from uterine tissues. They were characterized as Brucella abortus biovar 1 after biotyping by PCR, showing the fragment of 498 bp. Five of 8 farms were diagnosed as brucellosis two to four times more over the intervals of two or three months. Here in this study we briefly showed the correlation of the sporadic outbreak of brucellosis tested by SAT and the isolation of the causative organism. Moreover one or two reactors against brucellosis among considerable size of herd may indicate that SAT failed to detect potentially infected individuals in the incubation stage or chronic phase of the disease.

회전 원추형 마늘 쪽분리기 개발에 관한 연구 (II) - 원추 간극의 영향 - (Development of Rotating Cone Type Garlic Clove Separator (II) - Effect by Clearance between Inner and Outer Frustum -)

  • 이종수;김기복
    • Journal of Biosystems Engineering
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    • 제32권2호
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    • pp.77-83
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    • 2007
  • The purpose of this study is to find optimal conditions for various outlet clearances of prototype garlic clove separator with a rotating cone in the constant inlet clearance and cone height. Optimal outlet clearance from medium to small size garlics was 25 mm at the $200{\sim}400rpm$. For large garlic, optimal outlet clearances of Namdo and Uiseong garlic were 34 mm and 37 mm, respectively, in the range of $300{\sim}400rpm$. The proportion of garlic separation was over 95% for all quality of garlics. The proportions of damaged garlics at 25 mm and 28 mm outlet clearances were below 5% and below 10%, respectively. Therefore, in order to maintain high performance of garlic separation for the various varieties and qualities, the rotating cone type separator should be designed with cone speed ranges of $200{\sim}400rpm$ and the outlet clearance ranges of $25{\sim}37 mm$. The outlet clearance of the separator should be easily controlled within those ranges.

nif-Gene Organization and Nucleotide Sequence of nifV, nifH, D, K and nifE from Frankia Strain FaCl

  • An, Chung-Sun
    • 한국동물학회:학술대회논문집
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    • 한국동물학회 1995년도 한국생물과학협회 학술발표대회
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    • pp.120-120
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    • 1995
  • The total size of the pF AR1, a genomic clone of Frankia FaCI, was estimated to be about 44Kb by summation of the individual fragment length generated by single or double restriction enzymes. Southern hybridization analyses with Azotobacter vinelandii nif-genes as probes and partial sequencing analyses of the subclones revealed that organization of the nif-gene in the FaCI strain was nifV, H, D, K, E, N, X, W, B. The organization of the structural genes for nitrogenase is the same in this Frankia strain as it is in most other nitrogen-fixing prokaryotes but the positioning of the nifV-like gene relative to the nifHDK cluster differs. A consensus nif-promoter-like sequence, found at 5' of nifH, was not detected upstream of the niJV-like gene. nifV-like gene contained a ORF of 1206 NT encoding 401 amino acids. The nucleotide sequence and deduced amino acid sequence of the gene exhibit homology value of 65% and 41% with that from A vinelandii, respectively. The putative Shine-Dargamo sequences were present preceding nitK, nifH, D, K, and nifE, and in nitK gene putative start codon GTG was detected instead of A TG. The nucleotide and amino acid sequence of niIK of FaCI showed 82% and 76% homolgy with those of Frankia HFPCc 13, respectively. Amino acid sequence of niIK showed 69% and 61% homology with those of A vinelandii, Klebsiella pnewnoniae, respectively, while that of nifE 73% and 71%, respecti vely.i vely.

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Novel Trimeric Complex for Efficient Uptake of Plasmid Vector into HepG2 Cells

  • Joo, Jong-Hyuck;Park, Jong-Gu
    • 대한의생명과학회지
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    • 제9권2호
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    • pp.67-74
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    • 2003
  • Viral and non-viral vectors have been used in the delivery of genetic materials into animal cells and tissues, with each approach having pros and cons. Non-viral vectors have many useful merits such as easy preparation, low immunity and size tolerance of a transgene when compared to those of viral vectors. Delivery specificity may be achieved by complex formation between receptor ligands and a non-viral vector. In the present study, non-viral vector systems are investigated in an effort to find a practical delivery means for gene therapy, Receptor-ligand interaction between transferrin-receptor and transferrin was utilized for efficient gene transfer into cancer cells. A plasmid vector, pcDNA3 (LacZ) was ligated with a small duplexed oligo fragment in which a Biotin- VN$^{TM}$ phosphoramidite was placed in the middle of the oligo. The plasmid vector labeled by biotin was then conjugated with biotin-labeled transferrin via streptavidin. This trimeric conjugates were delivered to a hepatoma cell line, HepG2. The delivery efficiency of the trimeric conjugate was 2-fold higher than that of cationic liposomes used for transfection of a plasmid vector. These results demonstrate that a plasmid vector can be efficiently transferred into cells by forming a trimeric complex of plasmid vector-linker-ligand.

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악안면부 총상 환자의 전산화단층사진상의 연구 (COMPUTED TOMOGRAPHIC STUDY OF MAXILLOFACIAL GUNSHOT INJURIES)

  • 박인우
    • 치과방사선
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    • 제26권2호
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    • pp.65-73
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    • 1996
  • The purpose of this study was to evaluate the clinical and computed tomographic features of 7 cases of maxillofacial gunshot injuries in the suicidal patients visited the emergency room, Capital Armed Forces General Hospital. The obtained results were as follows : 1. The gunshot wounds were directed from submental area to dorsum of nose(3 cases), frontal area(1 case), orbit(1 case), infraorbital area (1 case), and lateral to nasal wing(l case). The shape of inlet in gunshot wounds were round (diameter: l-3cm) and that of outlet were oval shape(size : inlet

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두 가지 발파 패턴의 파쇄 성과 비교 (Comparison of Fragmentation Performance of Two Different Blast Patterns)

  • 피유시 라이;양형식
    • 터널과지하공간
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    • 제20권5호
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    • pp.325-331
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    • 2010
  • 이 연구에서는 사암 벤치에 대하여 V형과 경사 V형의 두 가지 기폭 패턴으로 대규모 발파를 실시하였다. 장약량은 비슷하였다. 발파 후 디지털 이미지 프로세싱 방법으로 파쇄석의 파쇄도를 평가하였다. 10 $m^3$ 용량 로우프 쇼벨의 싸이클 시간도 계측하였다. 그 결과는 경사 V형으로 기폭한 경우 파쇄성능이나 굴착기 성과가 더 향상된다는 것을 보여준다. 이 논문에서는 경사 V형 패턴이 V 형 패턴보다 더 나은 성능을 보이는 이유에 대하여 논한다.

병원성장내세균에서 phoP-phoQ operon의 지배를 받는 phoA 유전자의 cloning 및 염기서열결정 (Cloning and Sequencing of the phoA Gene which is Regulated by the phoP-phoQ Operon in Pathogenic Enteric Bacteria)

  • 김성광;이태윤
    • Journal of Yeungnam Medical Science
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    • 제12권2호
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    • pp.237-245
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    • 1995
  • Klebsiella pneumoniae 의 phoA 유전자를 함유하는 DNA를 plasmid pACYC184에 클로닝 하였다. 클로닝된 DNA의 크기는 4.0 kb이었으며 제한효소지도를 작성한 결과 3개의 PstI 절단부위와 4개의 PvuII 절단부위가 발견되었다. Klebsiella pneumoniae 의 phoA 유전자의 염기서열은 Escherichia coli와 매우 유사하여 80%의 유사성을 보였으며 이는 이 두 균종이 서로 유전적으로 매우 가까운 관계에 있음을 시사하였다.

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