• 대한소아치과학회지
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• 제25권2호
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• pp.292-303
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• 1998

The arbitrary primer polymerase chain reaction(AP-PCR) and Southern blot restriction fragment length polymorphism(RFLP) were used to genotype the cariogenic pathogen S. mutans in children. Following the morphologic chracteristics of colony on selective medium for S. mutans, total genomic DNA from 155 strains was extracted by conventional methods. Among 155 strains, 143 strains (92.3%) were confirmed S. mutans by PCR with dexA gene and 114 strains were used in this study. Three random sequence 10-base oligonucleotide primers were chosen for AP-PCR. The amplified DNA products were separated electrophoretically in a 2% agarose gel containing ethidium bromide and the banding patterns were compared among different strains. For RFLP analysis, DNA was digested with EcoRI and BamHI, separated on a 0.7 % agarose gel and transferred to a nylon membrane. The membrane was probed with a previously characterised 1.6 kilobases (kb) DNA fragment cloned from gtf B gene of S. mutans. The probe was labeled with isotope[$^{32}P-{\alpha}CTP$], and hybridized fragments were detected with intensifying screen. AP-PCR produced 4-8 DNA bands in the 0.25-10 kb regions and distinguished 9, 10 or 12 genotypes, depending on the specific primer used. Southern blot RFLP analysis revealed 2 hybridization patterns consisting of 1 DNA fragments 450, 500 bp. These results indicate that AP-PCR is more discriminative method for genotyping of S. mutans.

• 한국자원식물학회지
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• 제28권3호
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• pp.341-349
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• 2015

Magnoliae Flos (Sini in Korean) is one of the most important oriental medicinal plants. In the Korean Herbal Pharmacopeia, the bud of the all species in Manolia denudate and Manolia genus were regarded as the botanical sources for ‘Sini’. Most the dried bud of Manolia denudata, Manolia biondii and Manolia sprengeri were used as ‘Xin-yi’ in China. Therefore, the purpose of this study was to determine and compare the ‘Magnolia’ species, four species including Manolia denudata, M. biondii, M. liliiflora and M. Kobus were analysis of sequencing data revealed DNA polymorphisms. The based on tRNA coding leucine/phenylalanine (trnL-F) and NADH-plastoquinone oxidoreductase subunit 5 (ndhF) sequences in chloroplast DNA. For the identification of ‘Magnolia’ species, polymerase chain reaction (PCR) analysis of chloroplast DNA regions such as ndhF have proven an appropriate method. A single nucleotide polymorphism (SNP) has been identified between genuine “Sini” and their fraudulent and misuse. Specific PCR primers were designed from this polymorphic site within the sequence data, and were used to detect true plants via multiplex PCR.

• Mycobiology
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• 제29권3호
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• pp.121-131
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• 2001

This study was carried out to identify the phylogenetic relationships among several caterpillar fungi by comparing the sequences of internal transcribed spacer regions(ITS1 and ITS2) and 5.8S ribosomal DNA(rDNA) repeat unit. The sequences of ITS1, ITS2, and the 5.8S rDNA from 10 strains of Cordyceps species, 12 strains of Paecilomyces, 3 strains of Beauveria, 2 strains of Metarhizium and 1 strains of Hirsutella were amplified, determined and compared with the previously known Cordyceps species. The sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites could be found. In the phylogenetic tree, the species generally divided into three clusters, supported by their morphology and/or host ranges. The 5.8S rDNA and TTS1 sequences among 10 species of Cordyceps militaris were identical and only one base pair in ITS2 sequence was different. Cordyceps sinensis and Cordyceps ophioglossoides were also clearly different, although they belonged to the same cluster. The Geniank database search of species revealed sister taxa of an entomogenous fungus. Metarhizium was used as an putgroup in all taxa.

• 사상체질의학회지
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• 제21권2호
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• pp.144-152
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• 2009

1. Objective : This study aims to report symptomatic improvement of a Soyangin patient reporting general weakness, dizziness, gait disturbance, cold sensitivity, nocturnal sweating, and anorexia after treatment with Hyeongbangjihwang-tang. 2. Method : The patient's subjective and objective symptoms were observed daily, and the VAS scores for the main symptoms were recorded daily throughout the hospitalization period. 3. Result : The general condition of the patient improved by the end of the hospitalization period, where the VAS score dropped from VAS 10 to VAS 5 and the symptoms of dizziness, general weakness, cold sensitivity, and nocturnal sweating disappeared by day 4 of admission without recurrence. 4. Conclusion : Although the patient did not report diarrhea, one of the main Mangeum symptoms, the general symptoms improved with Hyeongbangjihwang-tang medication. This indicates that Hyeongbangjihwang-tang can be used to treat chief and subsidiary symptoms related to Mangeum symptomatic pattern when applied to the appropriate base and manifesting symptomology.

• 대한수의학회지
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• 제41권2호
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• pp.189-195
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• 2001

Single base substitution and deletion mutation have been introducted into the verotoxin 2 (VT2)A subunit gene from O157:H7 isolates to reduce cytotoxicity of VT2 and the cytotoxicity between wild type toxin and mutant toxoid were compared. A M13-derived recombinant plasmid pEP19RF containing a 940bp EcoRI-PstI fragment of VT2A gene was constructed for oligonucleotide-directed mutagenesis. The duoble mutant pDOEX was constructed by point and deletion mutation of two different highly conserved regions of VT2A encoding active site cleft of enzymatic domain. The key residue, Glu 167(GAA) and the pentamer(WGRIS) consisting of the enzymatic domain were replaced by ASP(GAC) and completely deleted in nucleotide sequence analysis of mutant, respectively. In the comparision of vero cell cytotoxicity between wide type toxin and toxoid from mutant, the wild type toxin expressed cytotoxicity in dilution of $10^{-6}$, but the toxid from mutant did not show cytotoxicity to vero cells.

• 한국잡초학회지
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• 제18권1호
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• pp.76-83
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• 1998

피속 잡초 수집종 33종을 대상으로 RAPD marker를 이용하여 피 수집종 간의 유전적변이를 알아보고, 수집종들을 판별할 수 있는 DNA marker를 선발하기 위하여 본 실험을 수행한 결과를 요약하면 다음과 같다. Operon사에서 제작된 74개의 10-mer RAPD primer 가운데에서 명확한 다형성을 보이는 6개 primer를 선발하였다. 이들 primer로 PCR에서 증폭된 밴드는 31개이었으며 이 가운데 다형성을 나타내는 band는 26개(83.9%)로 나타났다. 피 수집종 간의 유연 관계를 분석한 결과 공시된 피 수집종은 크게 3그룹으로 분류할 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제22권6호
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• pp.742-753
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• 2012

Copper-containing compounds are introduced into the environment through agricultural chemicals, mining, and metal industries and cause severe detrimental effects on ecosystems. Certain microorganisms exposed to these stressors exhibit molecular mechanisms to maintain intracellular copper homeostasis and avoid toxicity. We have previously reported that the soil bacterial isolate Achromobacter sp. AO22 is multi-heavy metal tolerant and exhibits a mer operon associated with a Tn21 type transposon. The present study reports that AO22 also hosts a unique cop locus encoding copper homeostasis determinants. The putative cop genes were amplified from the strain AO22 using degenerate primers based on reported cop and pco sequences, and a constructed 10,552 base pair contig (GenBank Accession No. GU929214). BLAST analyses of the sequence revealed a unique cop locus of 10 complete open reading frames, designated copSRABGOFCDK, with unusual separation of copCD from copAB. The promoter areas exhibit two putative cop boxes, and copRS appear to be transcribed divergently from other genes. The putative protein CopA may be a copper oxidase involved in export to the periplasm, CopB is likely extracytoplasmic, CopC may be periplasmic, CopD is cytoplasmic/inner membrane, CopF is a P-type ATPase, and CopG, CopO, and CopK are likely copper chaperones. CopA, B, C, and D exhibit several potential copper ligands and CopS and CopR exhibit features of two-component regulatory systems. Sequences flanking indicate the AO22 cop locus may be present within a genomic island. Achromobacter sp. strain AO22 is thus an ideal candidate for understanding copper homeostasis mechanisms and exploiting them for copper biosensor or biosorption systems.

• Journal of Microbiology and Biotechnology
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• 제20권12호
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• pp.1653-1663
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• 2010

The first gene (${\alpha}$-gal1) encoding an extracellular ${\alpha}$-Dgalactosidase from the thermophilic fungus Talaromyces emersonii was cloned and characterized. The ${\alpha}$-gal1 gene consisted of an open reading frame of 1,792 base pairs interrupted by six introns that encoded a mature protein of 452 amino acids, including a 24 amino acid secretory signal sequence. The translated protein had highest identity with other fungal ${\alpha}$-galactosidases belonging to glycosyl hydrolase family 27. The ${\alpha}$-gal1 gene was overexpressed as a secretory protein with an N-terminal histidine tag in the methylotrophic yeast Pichia pastoris. Recombinant ${\alpha}$-Gal1 was secreted into the culture medium as a monomeric glycoprotein with a maximal yield of 10.75 mg/l and purified to homogeneity using Hisbinding nickel-agarose affinity chromatography. The purified enzyme was maximally active at $70^{\circ}C$, pH 4.5, and lost no activity over 10 days at $50^{\circ}C$. ${\alpha}$-Gal1 followed Michaelis-Menten kinetics ($V_{max}\;of\;240.3{\mu}M/min/mg,\;K_m\;of\;0.294 mM$) and was inhibited competitively by galactose ($K_m{^{obs}}$ of 0.57 mM, $K_i$ of 2.77 mM). The recombinant T. emersonii ${\alpha}$-galactosidase displayed broad substrate preference, being active on both oligo- and polymeric substrates, yet had strict specificity for the ${\alpha}$-galactosidic linkage. Owing to its substrate preference and noteworthy stability, ${\alpha}$-Gal1 is of particular interest for possible biotechnological applications involving the processing of plant materials.

• 전자공학회논문지SC
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• 제49권1호
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• pp.20-28
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• 2012

보행을 관찰하고 객관적인 정보를 추출하여 그 기능을 평가하는 것을 보행 분석이라고 한다. 최근 사용되는 보행 측정 장비들은 다수의 카메라, 지면 반력 측정 장치로 구성되어 고가이며, 이를 설치할 넓은 장소를 필요로 한다는 단점이 있다. 본 연구에서는 이러한 문제점을 해결하기 위해 단일 카메라를 통해 얻은 영상에서 마커 없이 인체의 3차원 보행 동작을 측정하는 기법을 제안한다. 파티클 필터를 사용하여 훈련 데이터와 보행에 관한 사전 정보 없이 사람의 동작을 추적한다. 인체와 지면에 관한 동역학을 통해 물리적으로 합당한 인체의 동작들을 생성하였다. 보행 영상에서 계산한 모든 관절의 평균 에러는 제안한 방법에서 $12.4^{\circ}$로, 기존 파티클 필터의 에러 $34.6^{\circ}$보다 작았다. 이러한 결과를 바탕으로 단일 카메라만으로 보행을 정량적으로 측정하여 기존 복잡한 장비를 대체할 수 있는 가능성을 제시하였다.

• 한국동물학회지
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• 제39권1호
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• pp.54-64
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• 1996

Xenopus를 사용하여 후각원판이 정상적인 상태에서 어떠한 과정을 거쳐 발달하는지 연구하였다. 특히 뉴런의 형태적 분화, 초기 발달과 성숙 양상, 축색과 원시전뇌의 접촉 등에 초점을 맞추었다. 후각원판은 stage 23에 외배엽이 두터워진 형태로 처음 나타나는데, 쌍을 이루며 각각은 표피측에 비신경층(NNL)과 안쪽의 신경층(NL)의 두 층으로 되어 있다. stage 26 후에 원판 세포는 NNL세포 틈을 비집고 상피쪽으로 이동하기 시작하며, stage 28이 되면 선단 돌기가 표피 끝에 도달한다. stage 29/30에는 NL의 기부에서 기부 돌기(미래의 축색)가 나타나 stage 32무렵 종뇌에 도달한다. 시냅스는 stage 37/38에 처음 나타난다. 일부 원판 세포들이 후각 뉴런으로 분화하는 동안 많은 원판 세포들은 기저세포로서 후각상피에 그대로 남아 있다. 연구 결과는 뉴런 외배엽의 NL에서 기원하고 지지세포는 NNL층에서 기원함을 보여주었다. 또한 시냅스 형성 전에 뉴런의 분화가 완성됨으로써 후각뉴런의 분화는 뇌의 발달과 독립적으로 일어나며 뇌의 영향을 받지 않는다는 사실을 알 수 있었다.