• 생명과학회지
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• 제15권6호
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• pp.851-856
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• 2005

Although valuable microbes have been isolated from the soil for the various productions of useful components, the microbes which can be cultivated in the laboratory are only $0.1-1\%$ of all microbes. To solve this problem, the study has recently been tried for making the valuable components from the environment by directly separating unculturable micrbial DNA in the soil. But it is known that humic acid originated from the soil interrupts various restriction enzymes and molecular biological process. Thus, in order to prevent these problems, this study modified the method separated soil DNA with phenol, CTAB and PEG. In order to compare the degree of purity for each DNA and the molecular biological application process, $A_{260}/A_{280}$ ratio, restriction enzymes, and PCR were performed. In case of DNA by the modified method, total yield of DNA was lower but $A_{260}/A_{280}$ ratio was higher than the previously reported methods. It was confirmed that the degree of purity is improved by the modified method. But it was not cut off by all kinds of tested restriction enzymes because of the operation of a very small amount of interrupting substances. When PCR was operated with each diluted DNA in different concentrations and GAPDH primer, the DNA by the modified method could be processed for PCR in the concentration of 100 times higher than by the previously reported separation method. Therefore, this experiment can find out the possibility of utilization for the unknown substances by effectively removing the harmful materials including humic acid and help establishing metagenomic DNA library from the soil DNA having the high degree of purity.

• 한국문헌정보학회지
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• 제40권1호
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• pp.13-37
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• 2006

본 연구는 근거이론을 적용하여 임신부들의 정보추구경험과 관련된 제 개념 및 이들 개념들 간의 관계 규명을 통하여 임신부의 정보추구에 관한 구조와 과정을 밝힌 연구이다. 이론적 표집에 의해 16명의 임신부들을 대상으로 심층적인 면담을 수행하였고. 면담데이타는 근거이론의 지속적인 비교분석에 의해 분석되었다. 본 연구에서 임신부의 정보추구의 발생원인은 임신의 수용으로 나타났다. 임신부들은 지각된 불안과 궁금증의 맥락에서 임신의 정상상태 유지현상을 해결하기 위해 산전 진단데이타 추구하기. 표준지식 추구하기, 경험담 추구하기와 정서적 지지 추구하기의 작용/상호작용적 정보추구 전략을 취하였으며, 비교하기, 맥락화 하기, 확인하기의 정보추구의 과정들을 거쳐 갔다. 이때, 정보 획득방법 인지, 임신 출산의 선행지식, 자아조절, 정보접근환경이 중재조건으로 작용하였다. 그 결과 충족과 불충족을 경험하였다. 또한 임신부들은 임신의 과정동안 정보요구의 변화 발전에 따라, 수용국면, 적응국면, 구체국면, 전환국면을 통과하였다. 후속연구에서는 본 연구에서 밝혀진 정보추구의 구조와 과정을 통합하여 임신부의 정보추구행위를 총체적으로 설명하는 실체이론을 개발해야 할 것이다.

• 한국문헌정보학회지
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• 제47권3호
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• pp.227-250
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• 2013

"정사공신녹권(定社功臣錄券)"은 태종이 1398년 제1차 왕자의 난을 평정하는 데에 힘쓴 신하들을 정사공신으로 책봉한 후 반사(頒賜)한 문서이다. 이 연구는 현존하는 "정사공신녹권" 복제본을 대상으로, 녹권이 반사된 사유 및 경위를 알아보았다. 녹권의 형태와 그 체제를 자세히 분석하여, 이를 바탕으로 녹권에 기재된 공신들을 각 등급별로 나누어 책봉된 공신들의 직함을 분석하여 특성을 찾고자 하였다. 그리고 녹권의 내용을 세세히 분석한 연구결과, "정사공신녹권"은 필사본으로 크게 권수(卷首) 본문(本文) 권말(卷末)의 3부분의 체계로 구성되었다. 또한 모두 29명(1등 12명, 2등 17명)이 기재되었으며, 그들의 직함과 성명이 기술되었다. 1등공신 직함의 품계가 2등에 비해 높았고 비교적 왕의 친인척이 많았다. 그리고 이들에 대한 포상 및 특전규정이 등급에 따라 다름을 확인하였으며, 같은 등급 안에서도 사람에 따라 그 규정 내용이 상이(相異)하였다. 공신에 관한 제반사무를 맡아본 공신도감의 관원들의 구성과 녹권 안 그들의 성씨(姓氏)와 수결을 통해 관원들의 상세한 성명을 파악하였다.

• 한국양식학회지
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• 제17권1호
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• pp.1-7
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• 2004

A rainbow trout uncoupling protein 3 (UCP3) cDNA clone, encoding a 310 amino acid protein, was cloned and sequenced from a liver cDNA library. Two different splice variants designated UCP3-vl and UCP3-v2, were identified through liver cDNA library screening using rainbow trout UCP3 cDNA clone as a probe. UCP3-vl has 3 insertions in the UCP3 cDNA: the first insertion (133 bp), the second (141 bp), and the third (370 bp) were located 126 bp, 334 bp and 532 bp downstream from the start codon, respectively. UCP3-v2 contained a single insertion, identical in sequence and location to the second insertion of UCP3-vl. UCP3, a mitochondrial protein, functions to modulate the efficiency of oxidative phosphorylation. UCP3 has been detected from heart, testis, spinal cord, eye, retina, colon, muscle, brown adipose tissue and white adipose tissue in mammalian animals. Human and rodent UCP3s are highly expressed in skeletal muscle and brown adipose tissue, while they show weak expression of UCP3 in heart and white adipose tissue. In contrast to mammalian studies, RT-PCR and Southern blot analysis of the rainbow trout demonstrated that UCP3 is strongly expressed in liver and heart. UCP3, UCP3-vl, and UCP3-v2 all contain an Ava III short interspersed element (SINE), located in the 3'untraslated region (UTR). PCR using primers from the Ava III SINE and the UCP3 3'UTR region indicates that the UCP3 cDNA is structurally conserved among salmonids and that these primers may be useful for salmonid species genotyping.

• 한국어류학회지
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• 제23권1호
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• pp.21-29
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• 2011

볼락의 성장단계에 따른 차등발현 유전자를 탐색하기 위하여 6개월령 및 18개월령 근육조직을 사용하여 subtracted cDNA library를 제작하였고, 각각의 연령에서 발현량 차이를 나타낸 202개의 cDNA 단편을 확보하였으며, 발현량 차이가 뚜렷한 32개의 cDNA 클론은 성장단계별 특이발현 후 보유전자로 선발하여 염기서열을 분석하였다. Myosin, adenylate kinase, calsequestrin, dystrobrevin beta, diphosphate kinase 유전자는 6개월령 근육조직에서 발현량이 많았으며, desmin, TGFBR2 (transforming growth factor-beta receptor), creatine kinase (muscle type), cathepsin D 유전자는 18개월령 근육조직에서 발현량이 많았다. 볼락의 성장초기와 성장절정기에서 차등발현 양상을 나타낸 유전자는 6, 18, 30, 42개월령 근육조직에서 연령 증가에 따른 발현양상을 분석하였으며, dystrobrevin beta와 diphosphate kinase-Z1은 6개월령 이후에는 발현량이 급격히 감소하여 18개월령, 30개월령 및 42개월령에서는 발현량이 극히 적었으며, creatine kinase (muscle type)와 cathepsin D 유전자는 연령 이 증가함에 따라 발현량이 증가되어 18개월령 이후, 30개월령과 42개월령 근육조직에서도 발현량이 많았다. 이와 같이 성장단계에 따른 차등발현 유전자를 탐색하고 연령 증가에 따른 발현양상을 비교 분석한 결과로부터 본 연구에서는 어류의 성장 초기단계 근육조직에서는 근육수축 관련 유전자가 많이 발현되고, 성장 절정기에는 근육 내 에너지 양 조절 관련 유전자가 많이 발현되는 것을 확인하였다.

• 생명과학회지
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• 제27권4호
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• pp.398-407
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• 2017

BTG1 (B-cell translocation gene 1)은 APRO family (anti-proliferative protein family)에 속하며, 이들은 공통의 생물학적 기능은 세포증식을 억제하는 것으로 알려져 있다. 본 연구에서, 굴의 gill cDNA library를 random sequencing을 통한 EST 분석과정에서 BTG1 clone을 확보하였으며, 분자생물학적 특성을 조사하였다. 굴의 BTG1은 182 개의 아미노산으로 구성되며, zebrafish와 57%, human과 56%의 상동성을 나타냈으며, 사람이나 설치류와 달리 ORF (open reading frame) 내에 intron이 존재하지 않았다. Genomic DNA walking을 통해 굴의 BTG1의 predicted promoter를 확인하였으며, 분석결과 AP-1 element와 SRE (serum response element) 부위가 존재하였으며, 5'flanking region에 cAMP response element (CRE) 부위가 확인되었다. 굴의 BTG1의 조직별 유전자발현 수준을 확인하기 위해 real-time PCR을 수행하였으며, 6 개 조직 모두에서 BTG1의 유전자발현이 나타났으며, 그 중에서 heart와 mantle에서 높은 수준의 mRNA 발현을 확인할 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제17권8호
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• pp.1330-1337
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• 2007

To better understand the gene expression of the cold-adapted polar diatom, we conducted a survey of the Chaetoceros neogracile transcriptome by cDNA sequencing and expression of interested cDNAs from the Antarctic diatom. A non-normalized cDNA library was constructed from the C. neogracile, and a total of 2,500 cDNAs were sequenced to generate 1,881 high-quality expressed sequence tags (ESTs) (accession numbers EL620615-EL622495). Based on their clustering, we identified 154 unique clusters comprising 342 ESTs. The remaining 1,540 ESTs did not cluster. The number of unique genes identified in the data set is thus estimated to be 1,694. Taking advantage of various tools and databases, putative functions were assigned to 939 (55.4%) of these genes. Of the remaining 540 (31.9%) unknown sequences, 215 (12.7%) appeared to be C. neogracile-specific since they lacked any significant sequence similarity to any sequence available in the public databases. C. neogracile consisted of a relatively high percentage of genes involved in metabolism, genetic information processing, cellular processes, defense or stress resistance, photosynthesis, structure, and signal transduction. From the ESTs, the expression of these putative C. neogracile genes was investigated: fucoxanthin chlorophyll (chl) a,c-binding protein (FCP), ascorbate peroxidase (ASP), and heat-shock protein 90 (HSP90). The abundance of ASP and HSP90 changed substantially in response to different culture conditions, indicating the possible regulation of these genes in C. neogracile.

• Asian-Australasian Journal of Animal Sciences
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• 제17권10호
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• pp.1329-1333
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• 2004

To isolate the microsatellites from the chromosomal DNA of the Korean cattle (Hanwoo) and to use those for the genetic selection, four bacteriophage genomic libraries containing the chromosomal DNA of six Hanwoo steers showing the differences in meat quality and quantity were used. Screening of the genomic libraries using $^{32}P-radiolabeled 5'-({CA})_{12}-3$nucleotide as a probe, resulted in isolation of about 3,000 positive candidate bacteriophage clones that contain $(CA)_n$-type dinucleotide microsatellites. After confirming the presence of microsatellite in each positive candidate clone by Southern blot analysis, the DNA fragments that include microsatellite and flanking sequences possessing less than 2 kb in size, were subcloned into plasmid vector. Results from the analysis of microsatellite length polymorphism, using twenty-two PCR primers designed from flanking region of each microsatellite DNA, demonstrated that 208 and 210 alleles of HW-YU-MS#3 were closely related to the economic traits such as marbling score, daily gain, backfat thickness and M. longissimus dorsi area in Hanwoo. Interestingly, HW-YU-MS#3 microsatellite was localized in bovine chromosome 17 on which QTLs related to regulation of the body fat content and muscle ypertrophy locus are previously known to exist. Taken together, the results from the present study suggest the possible use of the two alleles as a DNA marker related to economic trait to select the Hanwoo in the future.

• IMMUNE NETWORK
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• 제13권4호
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• pp.133-140
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• 2013

Since the discovery of the immunomodulation property of mesenchymal stem cells (MSCs) about a decade ago, it has been extensively investigated whether MSCs can be used for the treatment of immune-related diseases, such as graft versus-host disease (GvHD). However, how to evaluate the efficacy of human MSCs for the clinical trial is still unclear. We used an MHC-mismatched model of GvHD (B6 into BALB/c). Surprisingly, the administration of the human MSCs (hMSCs) could reduce the GvHD-related mortality of the mouse recipients and xenogeneically inhibit mouse T-cell proliferation and $IFN-{\gamma}$ production in vitro. We recently established a new protocol for the isolation of a homogeneous population of MSCs called subfractionation culturing methods (SCM), and established a library of clonal MSC lines. Therefore, we also investigated whether MSCs isolated by the conventional gradient centrifugation method (GCM) and SCM show different efficacy in vivo. Intriguingly, clonal hMSCs (hcMSCs) isolated by SCM showed better efficacy than hMSCs isolated by GCM. Based on these results, the MHC-mismatched model of GvHD may be useful for evaluating the efficacy of human MSCs before the clinical trial. The results of this study suggest that different MSC lines may show different efficacy in vivo and in vitro.

• Molecules and Cells
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• 제23권1호
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• pp.108-114
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• 2007

The mitogen-activated protein kinase (MAPK) signaling cascade is critical for regulating plant defense systems against various kinds of pathogen and environmental stresses. One component of this cascade, the MAP kinase kinases (MAPKK), has not yet been shown to be induced in plants following biotic attacks, such as those by insects and fungi. We describe here a gene coding for a blast (Magnaporthe grisea)- and insect (Nilaparvata lugens)-responsive putative MAPK kinase, OmMKK1 (Oryza minuta MAPKK 1), which was identified in a library of O. minuta expressed sequence tags (ESTs). Two copies of OmMKK1 are present in the O. minuta genome. They encode a predicted protein with molecular mass 39 kDa and pI of 6.2. Transcript patterns following imbibition of plant hormones such as methyl jasmonic acid (MeJA), ethephone, salicylic acid (SA) and abscisic acid (ABA), as well as exposure to methyl viologen (MV), revealed that the expression of OmMKK1 is related to defense response signaling pathways. A comparative analysis of OmMKK1 and its O. sativa ortholog OsMKK1 showed that both were induced by stress-related hormones and biotic stresses, but that the kinetics of their responses differed despite their high amino acid sequence identity (96%).