• 한국미생물학회:학술대회논문집
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• 한국미생물학회 1991년도 춘계학술발표대회 논문집
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• pp.225-236
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• 1991

The genes encoding the thermostable $\alpha$-amylase and maltogenic amylase from Bacillus lichenciformis were cloned and expressed in E. coli. The recombinant plasmid pTA322 was found to contain a 3.1kb EcoRI genomic DNA fragment of the thermostable $\alpha$-amylase. The cloned $\alpha$-amylase was compared with the B. licheniformis native $\alpha$-amylase. Both $\alpha$-amylase have the same optimal temperature of $70^{\circ}C$ and are stable in the pH range of 6 and 9. The complete nucleotide sequences of the thermostable $\alpha$-amylase gene were determined. It was composed of one open reading rame of 1,536 bp. Start and stop codons are ATG and TAG. From the amino acid sequence deduced from the nucleotide sequence, the cloned thermostable $\alpha$-amylase is composed of 483 amino acid residues and its molecular weight is 55,200 daltons. The content of guanine and cytosine is $47.46mol\%$ and that of third base codon was $53_41mol\%$. The recombinant plasmid, pIJ322 encoding the maltogenic amylase contains a 3.5kb EcoRI-BamHI genomic DNA fragment. The optimal reaction temperature and pH of the maltogenci amylase were $50^{\circ}C$ and 7, respectively. The maltogenic amylase was capable of hydrolysing pullulan, starch and cyclodextrin to produce maltose from starch and panose from pullulan. The maltogenic amylase also showed the transferring activity. The maltogenic amylase gene is composed of one open reading frame of 1,734bp. Start and stop codons are ATG and ATG. At 2bp upstream from start codon, the nucleotide sequence AAAGGGGGAA seems to be the ribosome-binding site(RBS, Shine-Dalgarno sequence). A putative promoter(-35 and-10 regions) was found to be GTTAACA and TGATAAT. From deduced amino acid sequence from the nucleotide srquence, this enzyme was comosed of 578 amino acid residues and its molecular weight was 77,233 daltons. The content of guanine and cytosine was $48.1mol\%$. The new recombinant plasmid, pTMA322 constructed by inserting the thermostable $\alpha$-amylase gene in the EcoRI site of pIJ322 to produce both the thermostable $\alpha$-amylase and the maltogenic amylase were expressed in the E. coli. The two enzymes expressed from E. coli containing pTMA322 was reacted with the $15\%$ starch slurry at $40^{\circ}C$ for 24hours. The distribution of the branched oligosaccharides produced by the single-step process was of the ratio 50 : 50 between small oligosaccharide up DP3 and large oligosaccharide above DP3.

• 식물조직배양학회지
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• 제28권5호
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• pp.283-287
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• 2001

감자 (Solanum tuberosum L. cv. Irish Cobbler)의 괴경형성과정 (tuberization) 동안에 발현하는 유전자들의 발현양상을 조사하고자 differential display법을 실시하였다. Differential display를 이용하여 분리된 eIF5A DNA단편을 probe로 사용하여 감자의 cDNA library screening을 통하여 eIF5A full-length cDNA를 감자에서 처음으로 분리하였다. 감자의 eIF5A, clone은 토마토의 eIF5A cDNA 염기서열과 94.8%. 아미노산 서열에서는 97.5%로 매우 높은 유사성을 나타내었다. 감자의 eIF5A 유전자는 길이가 716 bp로 하나의 단백질 code영역 (ORF)을 포함하고 있었다. 이 영역은 분자량 17.4 kD, pI 5.5로 추정되는 160개의 아미노산으로 구성된 eIF5A단백질을 code하고 있었다. eIF5A 단백질들에서 12개의 아미노산 서열 (STSKTGKHGHAK)은 효모에서 사람에 이르기까지 완벽하게 보존되어 있는 것으로 알려져 있는데, 감자에서도 또한 잘 보존되어 있었다. 이 영역은 eIF5A 단백질의 활성을 나타내는 데 있어서 필수적인 hypusine을 생성하는 전사 후 수식 부위가 들어 있는 아주 중요한 곳이다. 감자에서 eIF5A 유전자의 발현양상을 조사한 결과 감자의 전조직에서 발현을 보였는데, 성숙잎이나 괴경보다는 세포분열 및 물질축적이 활발히 일어나고 있는 꽃기관들 (stamen, ovary, petal. sepal), 과실 (fruit)과 stolen 등의 조직들에서 비교적 활발히 발현되고 있었다.

• 생명과학회지
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• 제12권2호
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• pp.188-199
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• 2002

Xylanase를 생산하는 내열성 Bacillus pumilus TX703의 chromosomal DNA로부터 xylanase 유전자를 cloning하여 그 염기배열 순서를 결정한 다음 이로부터 유전자 발현에 관련된 구조를 분석하였다. Xylanase 유전자의 cloning을 위해 제한효소 HindIII로 절단한 B. pumilus TX703의 chromosomal DNA와 pUC19을 ligation시켜 E. coli DH5 $\alpha$에 형질전환시킨 후 형질전환체 중에서 xylanase 활성을 나타내는 재조합 plasmid pXES106을 분리하였다. 재조합 plasmid pXES106은 pUC19의 HindIII 부위 내에 2.24 kb의 외래 DNA가 삽입되었고, 이 plasmid DNA를 분리하여 E. coli DH5 $\alpha$에 재형질전환시킨 결과 vector 내에 xylanase 유전자가 cloning되었음을 확인하였다. Cloning된 유전자의 염기배열을 분석한 결과 이 유전자의 총 크기는 2,187 bp였고 이는 409개기 아미노산을 coding 하는 open reading frame 1,227 bp를 포함하고 있었다. 이 염기배열은 ATG개시 codon으로부터 각각 193과 216 base 상류에 TTTAAT의 -10 box와 TCGAAA인 -35 box로 추정되는 염기배열이 존재하였고 -10 box로부터 7 bp하류에 전사개시점인 A가 위치하고 있었다. 또한, 개시 codon으로부터 432 bp 상류에 공통염기배열과 14개의 염기 중 11개의 염기가 일치하는 TGATGGCGTCGGCA의 catabolite responsive element (CRE)가 존재하였다. B. pumilus TX703의 xylanase와 아미노산배열의 유사성이 가장 높은 xylanase는 Hordeum vulgare의 isozyme X-I이었고 본 xylanase는 208번째와 322번째에 glutamic acid 잔기를 가지고 있어 Clostridium thermocellum, Dictyoglomus thermophilum, Thermotoga neapolitana 등에서 밝혀진 바와 같이 glutamic acid 부위가 xylanase의 활성부위라 여겨진다.

• 한국초등수학교육학회지
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• 제8권1호
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• pp.1-21
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• 2004

이야기 틀(story shell)을 활용한 수학 수업에서 학생들이 보여주는 수학적 의사소통의 현상을 분석하고, 이를 바탕으로 이미 규범화되어 있는 수학 수업에서 이루어지는 의사소통 지도 개선에 시사하는 바를 얼고자 하였다. 이야기 틀이란, 수학과 관련 있는 상황을 설정하는 하나의 수업 테크닉이다. 이야기 틀을 활용한 수학 수업에서는 학생들이 제시된 문제를 듣거나 저을 때 자신의 의식을 집중함으로써 문제 해결의 단서를 더 쉽게 찾는다는 것을 일 수 있었다. 따라서 이야기 틀을 수학적 문제해결에서의 이해를 강화하는 수단으로 제안할 수 있다. 또한, 이야기 틀은 그 속에 학생들의 생활과 관련된 것이 담겨 있고, 여러 가지 문학적 요소로 인해 학생들의 수학적 언어 사용에 도움을 준다. 그리고 이야기로 제시되는 문제 상황은 학생들의 흥미를 자극하고 동기를 부여함으로써 수학에 대한 학생들의 태도를 긍정적으로 이끌어 내며, 자신의 의견을 제시할 때 논리적으로 설명하는 것을 볼 수 있었다.

• 생명과학회지
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• 제15권6호
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• pp.1005-1012
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• 2005

식물 내생균 Bacillus sp. CY22는 식물병원균 Rhizoctonia solani, Fusarium oxysporum 및 Phythium ultimum에 대해 강한 항균력을 나타내었다. 일반적으로 많은 Bacillus속 균주들은 iturin, fengycin, mycosubtulin과 같은 항균 물질을 분비한다. 본 연구에서는 식물내생균 Bacillus sp. CY22의 배양액으로부터 항균물질을 분리, 정제하였으며 MALDI-TOF mass로 분자량을 확인하였다. MALDI-TOF mass spectrum분석 결과 분리된 항균물질은 Bacillus 속 균주가 생성하는 항균물질로서 잘 알려져 있는 iturin의 분자량과 거의 일치하였으며, m/z 1043.4, 1057.4, 1071.4에서 molecular ion peak를 나타내었다. 이들은 각각 m/z 14차이를 가진 iturin의 isoform으로 추정되며 이 것은 iturin을 구성하고 있는 지방산의 탄소수 차이로 생각되며 m/z 1065.4, 1079.4 peak는 sodium adduct로서 추정된다. 또한 항균물질 iturin을 생성하는데 관여하는 transacylase 유전자를 크로닝하여 ita22 유전자로 명명하고, 그 특성으로 ita22 유전자는 400 개의 아미노산을 인지하는 1,200 bp의 open reading frame (ORF)을 가지며, 아미노산의 상동성을 조사한 결과 Bacillus subtilis 168의 FenF (BAB69697)와 가장 유사하였다.

• 보건교육건강증진학회지
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• 제2권1호
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• pp.3-50
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• 1984

The levels of health knowledge, attitude and practice of housewives considerably effect to the health of households, communities and the nation. This study was designed to grasp the levels of health knowledge, attitude and practice of houswives and analyse the various factors effecting to health in order to provide health education services as well as materials for effective formulation and implementation of health policy to improve the health of the nation. This study has been conducted through interviews by trained surveyers for 4,281 housewives selected from 4,500 households throughout the country for 40 days during July 11-August 20, 1983. The results of survey were analysed by stepwise multiple regression and path analysis are summarized as follows; 1. Based on the measurement instrument applied to this study, the levels of health knowledge, attitude and practice of housewives were extremely low with 54.5 points out of 100 points in full. Higher level with 72 points and above was approximately 21 percent and lower level with 39 points and below was approx. 24 percent. The middle level was approx. 55 percent. In order to implement health programs successively, health education should be more strengthened and to improve the level of health knowledge, attitude and practice (KAP) of the nation, political consideration as a part of spiritual reformation must be concentrated on health. 2. The level of health knowledge indicated the highest points with 57.3 the level of attitude was the second with 55.0 points and the practice level was the lowest with 50.0 point. Therefore, planning and implementation of health education program must be based on the persuasion and motivation that health knowledge turn into practice. 3. Housewives who had higher level of health knowledge, showed their practice level was relatively lower and those who had middle or low level of it practice level was the reverse. 4. Correlations among health knowledge, attitude and practice (KAP) were generally higher and statistically significant at 0.1 percent level. Correlation between total health KAP level and health knowledge was the highest with r=.8092. 5. Health KAP levels showed significant differences according to the age, number of children, marital status, self-assessed health status and concern on health of the housewives interviewed (p<0.001) 6. Health KAP levels also showed significant differences according to the education level, economic status, employment before marriage and grown-up area of the housewives interviewed. (p<0.001) 7. Heath KAP levels showed significant differences according to health insurance benificiary and the existence of patients in the family. (p<0.001). 8. Health KAP levels showed significant differences according to distance to government organizations, schools, distance to health facilities, telephone possession rate, television possession rate, newspaper reading rate and activities of Ban meeting and Women's club. (p<0.001) 9. Health KAP levels showed significant differences according to electric mass communication media such as television, radio and village broadcasting etc. and printed media such as newspaper, magazine and booklets etc., IEC variables such as individual consultation and husband-wife communication, however, there was no significance with group training. 10. Health KAP of the housewives showed close correlation with personal characteristics variables, i.e., education level (r=.5302), age (r=-.3694) grown-up area (r=.3357) and employment before marriage. In general, correlation of health knowledge level was higher than the levels of attitude or practice. In case of health concern and health insurance, correlation of practice level was higher than health knowledge level. 11. Health KAP levels showed higher correlation with community environmental characteristics, Ban meeting and activity of Women's club, however, no correlation with New-village movement. 12. Among IEC variables, husband-wife communication showed the highest correlation with health KAP levels and printed media, electric mas communication media and health consultation in order. Therefore, encouragement of husband-wife communication and development of training program for men should be included in health education program. 13. Mass media such as electric mass com. and printed media were effective for knowledge transmission and husband-wife communication and individual consultation were effective for health practice. Group training was significant for knowledge transmission, however, but not significant for attitude formation or turning to health practice. To improve health KAP levels, health knowledge should be transmitted via mass media and health consultation with health professionals and field health workers should be strengthened. 14. Correlation of health KAP levels showed that knowledge level was generally higher than that of practice and recognized that knowledge was not linked with attitude or practice. 15. The twenty-five variables effecting health KAP levels of housewives had 41 per cent explanation variances among which education level had great contribution (β=.2309) and electric mass com. media (β=.1778), husband-wife communication (β=.1482), printed media, grown-up area, and distance to government organizations in order. Variances explained (R²) of health KAP were 31%, 15%, and 30% respectively. 16. Principal variables contributed to health KAP were education level (β=.12320, β=.1465), electric mass comm. media (β=.1762, β=.1839), printed media, (β=.1383, β=.1420) husband-wife communication (β=.1004, β=.1067), grown-up area and distance to government organizations, in order. Since education level contributes greatly to health KAP of the housewives, health education including curriculum development in primary, middle and high schools must be emphasized and health science must be selected as one of the basic liberal arts subject in universities. 17. Variences explained of IEC variables to health KAP were 19% in total, 14% in knowledge, 9% in attitude, and 10% in health practice. Contributions of IEC variables to health KAP levels were printed media (β=.3882), electric mass comm media (β=.3165), husb-band wife com. (β=.2095,) and consultation on health (β=.0841) in order, however, group training showed negative effect (β=-.0402). National fund must be invested for the development of Health Program through mass media such as TV and radio etc. and for printed materials such as newspaper, magazines, phamplet etc. needed for transmission of health knowledge. 18. Variables contributed to health KAP levels through IEC variables with indirect effects were education level (Ind E=0.0410), health concern (Ind E=.0161), newspaper reading rate (Ind E=.0137), TV possession rate and activity of Ban meeting in order, however, health facility showed negative effect (Ind E=-.0232) and other variables showed direct effect but not indirect effect. 19. Among the variables effecting health KAP level, education level showed the highest in total effect (TE=.2693) then IEC (TE=.1972), grown-up city (TE=.1237), newspaper reading rate (TE=.1020), distance to government organization (TE=.095) in order. 20. Variables indicating indirect effects to health KAP levels were; at knowledge level with R²=30%, education level (Ind E=.0344), newspaper reading rate (Ind E=.0112), TV possession rate (Ind E=.0689), activity of Ban meeting (Ind E=.0079) in order and at attitude level with R²=13%, education level (Ind E=. 0338), activity of Ban meeting (Ind E=.0079), and at practice level with R²=29%. education level (Ind E=.0268), health facility (Ind E=.0830) and concern on health (Ind E=.0105). 21. Total effect to health KAP levels and IEC by variable characteristics, personal characteristics variables indicated larger than community characteristics variables. 22. Multiple Correlation Coefficient (MCC) expressed by the Personal Characteristic Variable was .5049 and explained approximately 25% of variances. MCC expressed by total Community environment variable was .4283 and explained approx. 18% of variances. MCC expressed by IEC Variables was .4380 and explained approx. 19% of variances. The most important variable effected to health KAP levels was personal characteristic and then IEC variable, Community Environment variable in order. When the IEC effected with personal characteristic or community characteristic, the MCC or the variances were relatively higher than effecting alone. Therefore it was identified that the IEC was one of the important intermediate variable.

• 생명과학회지
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• 제28권11호
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• pp.1301-1315
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• 2018

참담치(Mytilus coruscus)의 혈구 유래의 항균 펩타이드를 역상 column들을 사용한 reversed-phase high-performance liquid chromatography (RP-HPLC)로 분리 및 정제하였다. 정제된 펩타이드는 matrix-assisted laser desorption ionization time-of-flight mass spectrophotometer (MALDI-TOF/MS)를 통해 분자량이 4041.866 Da으로 밝혀졌으며 Edman degradation법을 통해 25개의 N-말단 서열을 확보하였다. 이는 참담치의 mytilin B precursor와 100%, mytilin 8 precursor, mytilin 4 precursor와 96% 일치하였다. 또한 103개의 아미노산 서열을 코딩하고 있는 312 bp의 open-reading frame (ORF)을 밝혔으며 이는 참담치의 mytilin B precursor와 100% 일치하였다. 밝혀진 분자량과 아미노산 서열을 바탕으로 C-말단 alanine 잔기의 유무에 따라 2개의 펩타이드를 합성하였으며 이는 mytilin B1과 B2라고 명명하였다. 이들은 그람 양성 균주 Bacillus cereus, Streptococcus parauberis [minimal effective concentrations, MECs $41.6-89.7{\mu}g/ml$], 그람 음성 균주 Enterobacter cloacae, Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Providencia stuartii, Pseudomonas aeruginosa, Vibrio ichthyoenteri [MECs $7.4-39.5{\mu}g/ml$] 그리고 진균류인 Candida albicans [MECs $26.0-31.8{\mu}g/ml$]에 항균활성을 나타냈다. 본 연구 결과, 참담치 혈구 유래 mytilin B1과 mytilin B2는 넓은 항균 스펙트럼을 가지고 열과 염분에 대한 안정성이 높으며 용혈현상과 세포독성은 나타나지 않았다. 이러한 특성은 기능성 사료첨가제 및 항생제 대체제로써 충분히 안정적인 역할을 할 뿐만 아니라 추후 mytilin의 구조적 중요성과 참담치의 면역학적 측면에서 다양한 자료를 제시할 것으로 사료된다.

• 미생물학회지
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• 제40권2호
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• pp.81-86
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• 2004

Streptomyces coelicolor의 전 유전체 청보를 분석한 결과(7), 유전자 ID1103135가 코드 하는 open reading frame SCO7697이 phytase[myo-inositol hexakisphosphate phosphohydrolase상동성 (3-6,8,23)]에 유의하게 유사한 것으로 판단되었다. S. coelicolor A3(2)M의 염색체 DNA를 주형으로 PCR 방법으로 SCO7697 전체를 포함하는 DNA 단편을 클로닝하였다. 두 가지의 서로 다른 길이를 갖는 클로닝 된 ID1103135 DNA 단편을 E. coli 발현용 벡터pET728a(+)에 삽입하여,두 종의 재조합 벡터 pET28-SP와 pET28-LP를 얻었다. pET28-SP 와 pET28-LP를 각각 E. coli BL2l에 도입하여, IPTG로 발현 유도된 단백질을 SDS-polyacrylamide 전기영동으로 확인한 결과, 발현은 성공적으로 이루어 졌으나 대부분불용체를 형성하고 분자량은 예상보다 약간 큰 것으로 나타났다. 불용체 형성은 단백질의 불활성화를 수반 함으로서, 배양 온도를 $37^{\circ}C$에서 $30^{\circ}C$로 변화시켜 배양하는 방법으로 발현된 단백질을 가용화 시켰다. 발현된 단백질을 추출하여 조추출물 또는 정제한 상태로 phytase활성을 측청하였으나 효소활성은 관찰할 수 없었다. 대장균 시스템에서의 발현이 효소 활성의 소실을 초래했을 가능성이 있으므로, ID1103135 유전자를 자신의 promoter를 함유하도록 PCR 클로닝하여, E. coli - Streptomyces의 shuttle vector인 pWHM3에 삽입하고, 이를 방선균 호스트인 S. lividans에 도입하였다. 형질 전환체의 세포조추출액 및 세포배양액의 phytase 활성을 측청하였으나, 역시 활성을 확인할수 없었다. 이와 같은 결과는 SCO7697이 아주 높은 확률(E value; $6e^{-89}$)로 phytase일 것으로 annotation 되었으나, 실제는 이와는 다른 기능을 함유하고 있음을 시사하고 있다.

• 한국잠사곤충학회지
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• 제52권1호
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• pp.45-51
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• 2014

면역 유도된 천잠(Antheraea yamamai) 유충에서 cecropin-A 유전자를 분리하였고 이 유전자를 Ay-CecA로 명명하였다. 전체 Ay-CecA cDNA 크기는 419 bp로 64개의 아미노산 잔기를 인코딩하는 195 bp ORF로 구성되어 있다. 천잠 CecA 유전자는 22개 잔기의 signal peptide, 4개 잔기의 propeptide 및 항균활성을 갖는 37개 잔기로 구성된 성숙 단백질(mature protein) 영역으로 구성되고 예상 분자량은 4046.81 Da으로 산출되었다. 천잠 CecA의 아미노산 서열은 다른 나비목 곤충에서 분리된 cecropin와 매우 높은 상동성(62 ~ 78%)을 나타냈다. Ay-CecA 유전자의 C말단에 기존에 보고된 곤충의 cecropin에서와 동일하게 C말단 아미드화를 위한 glycine 잔기가 존재하고 있다. 이 펩타이드의 항균활성을 검정하기 위해서 대장균 발현 시스템을 이용하여 활성이 있는 재조합 Ay-CecA 발현에 성공하였다. 발현 기주인 대장균에 대한 재조합 CecA 독성 중화를 위해서 불용성 단백질인 ketosteroid isomerase(KSI) 유전자를 CecA 유전자와 융합하였다. 융합 CecA-KSI 단백질은 대부분 불용성 단백질로 발현되었다. 발현된 융합단백질은 Ni-NTA immobilized metal affinity chromatography(IMAC)에 의해서 정제하였으며 CNBr 반응을 통하여 재조합 CecA를 절단하여 용출하였다. 최종적으로 양이온 교환 chromatography 과정을 통하여 CecA를 순수 정제하였다. 정제된 재조합 Ay-CecA는 그람음성균인 E. cori ML 35, Klebsiella pneumonia 및 Pseudomonas aeruginosa에 대해 매우 높은 항균활성을 나타냈었다. 따라서 본 연구 결과, 높은 항균활성 지닌 CecA는 천잠의 면역 반응에서 중요한 역할을 담당할 것으로 사료된다.

• 한국도서관정보학회지
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• 제9권
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• pp.63-83
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• 1982

Among many open libraries under the rule of the Japanese imperialism, only the railroad library as the character of special library under the control of the Japanese Government General is remarkable for having opened its facilities to outsiders as well as the interior users. This study tries a) To look into what were the colonial background, aim and motive of the establishment of the railroad library under the Japanese rule, b) To analyze its activities and the change of its organization, c) And to find out its vestiges an the effects which it has on our country's library circles. The following are the result: A. The railroad library under the rule of the Japanese imperialism functioned as a public library by the outside book's circulation rather than the interior using. B. The outsiders were qualified to use it only when they got the joint guarantee for the city resident, which means that the o n.0, pportunity of the circulation was not free for the Koreans rather than the Japanese. C. From the aspect of collections, technical books such as natural science, and reference ones tended to be collected, and their whole composition was focused on the materials for the Japan-Korea identification scheme and development of the national spirit as the Japanese empire's subjects. D. In the respect of its organization, all the clerical employees, as well as the chief librarian were composed of the Japanese. Although the Koreans were in principle prohibited from working there, a few of them were employed as guard or servant as the China-Japan War was close to the end. E. The area of the outside book's, circulation service was expanded throughout the whole region along the railroad connecting Pusan with Manchuria. Such a service activity which contained several meanings, was tried by means of culture's books, circuit's books, train's books, hotel's books, mind's development books and reader's reading books etc. F. Such matters as the limitation on book circulating and the quantity of circulation book, the paid circulation and the partial restriction on free circulation mean the wartime system under the rule of the Japanese imperialism and also a n.0, ppeared as a factor of hampering the Koreans from getting accustomed to using library, which has affected them until now. G. The noticeable thing in the service activity is shown in attempting the diversified and positive way of service like free circulation in order to solidify the attitude of the Koreans toward war by organizing mind's development books as well as culture's books with the China-Japan War. H. On the other hand, it can not be passed over that the diversified service activity of the railroad library played a role of bridge in turning of today's library circles of ours to the modernization.