• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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• pp.14-17
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• 2001

Positional clonging (map-based cloning) of mutations or genetic variations has been served as an invaluable tool to understand in-vivo functions of genes and to identify molecular components underlying phenotypes of interest. Mice homozygous for the cerebellar deficient folia (cdf) mutation are ataxic, with cerebellar hypoplasia and abnormal lobulation of the cerebellum. In the cdf mutant cerebellum approximately 40% of Purkinje cells are ectopically located within the white matter and the inner granule cell layer (IGL). To identify the cdf gene, a high-resolution genetic map for the cdf-gene-encompassing region was constructed using 1997 F2 mice generated from C3H/HeSnJ-cdf/cdf and CAST/Ei intercross. The cdf gene showed complete linkage disequilibrium with three tightly linked markers D6Mit208, D6Mit359, and D6Mit225. A contig using YAC, BAC, and P1 clones was constructed for the cdf critical region to identify the gene. A deletion in the cdf critical region on chromosome 6 that removes approximately 150 kb of DNA selection. cdf mutant mice with the transgenic copy of the identified gene restored the brain abnormalities of the mutant mice. The positional cloning of cdf gene provides a good example showing the identification of a gene could lead to finding a new component of important molecular pathways.

• 미생물학회지
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• 제28권1호
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• pp.35-40
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• 1990

Thiostrepton 내성 유전자(tsr)를 포함하는 multi-copy 재조합 플라스미드 pJY7J2의 제한효소 절단지도를 작성하였다. pJY, 712는 Streptomyces에서 넓은 host range를 나타내었으며 cloning 목적에 사용할 수 있는 단일 BgtIl 제한효소 인식부위를 갖고 있었다. 플라스미드 pJY 712는 lethal zygosis(Ltz+) 현상을 보였다. pJY 712의 혁질전환빈도는 S. lividans에서 $5.0\times 10^{4}$ TFU였다. pJY 712의 Bell 제한효소 인식부위에 tyrosmase 유전자(mel)를 삽입하여 플라스미드 PJY713을 제조하였다. met 유전자를 포함한 재조합 플라스미드 pJY 714는 pJY 713의 일부분(1.9kb BgllI-BelI 단편)을 제거하여 제고하였다.

• 한국정보통신학회논문지
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• 제6권3호
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• pp.452-458
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• 2002

클라이언트에서 검색한 지도를 스크롤할 경우 이미 전송 받은 객체를 캐슁하여 무선 데이터 전송량을 줄일 수 있지만, 클라이언트 내에서 데이터 좌표 변환, 선택적인 객체 삭제, 압축 단계가 부가적으로 요구된다. 특히 좌표 변환 연산은 모바일 휴대폰의 낮은 계산 능력, 적은 메모리 용량 등과 같은 제한적 자원으로 많은 시간이 요구된다. 따라서 휴대폰 기반 모바일 지도 서비스에 서 효과적인 지도 제어 연산을 수행하기 위해서는 휴대폰의 제한적인 자원을 극복함과 통시에 무선 네트웍 대역폭을 줄이는 방안이 연구되어야 한다. 이 논문에서는 클라이언트-서버 시스템에서 이미 전송 받은 객체의 효율적인 캐슁 기법을 제안하고자 한다. 또 캐슁된 데이터의 재이용과 원시 데이터의 재전송 방법의 장단점에 분석하고자 한다.

• 대한화장품학회:학술대회논문집
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• 대한화장품학회 2003년도 IFSCC Conference Proceeding Book I
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• pp.285-290
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• 2003

Living cells are continuously subject to all sorts of stress such as ultraviolet rays on skin cells. Tests made in various laboratories show that when young fibroblasts (Le. at the beginning of their proliferate life) were repeatedly put under stress at subletal doses, they acquired a phenotype similar to Senescence Induced Prematurely by Stress (SIPS). The work presented hereafter was made on a new model of senescence induced prematurely by stress from ultraviolet Brays (UVB). The human fibroblast model was put under repeated UVB stress, causing SIPS. Several ageing biomarkers were used in order to characterise the cells that underwent stress:. an increase in the proportion of positive cells with senescence associated $\beta$-galactosidase activity (SA $\beta$-gal) measured by a specific coloration,. the proportion in the different morphological stages that fibroblasts undergo during culture visualised by microscopic observation,. the expression of genes known for overexpressing during senescence, particularly fibronectin and apolipoprotein J, measured by Real Time-PCR,. the common deletion of 4,977 bp in mitochondrial DNA, evaluated by nested PCR. Studying the variation of these 4 biomarkers, we have evaluated the protective effect of a Laminaria digitata extract (LDE) that can be used as a natural active ingredient for anti-ageing cosmetics.

• 한국가축번식학회지
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• 제26권4호
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• pp.395-399
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• 2002

Human chorionic gonadotropin (hCG) is a member of the glycoprotein hormone family which includes FSH. hCG TSH. These hormone family is characterized by a heterodimeric structure composed a common $\alpha$-subunit noncovalently linked to a hormone specific $\beta$-subunit. To determine u and $\beta$ -subunits can be synthesized as a single polypeptide chain (tethered-hCG) and also display biological activity, the tethered-hCC and -FSH molecule by fusing the carboxyl terminus of the hCG $\beta$-subunit to the amino terminus of the $\alpha$-subunit was constructed. To determine the importance of $\alpha$ COOH -terminal amino acid, we also deleted the $\alpha$ COOH-terminal amino acids. The expressing vectors were transfected into CHO-K 1 cells. The tethered-wthCG and -wtFSH was efficiently secreted. The $\alpha$ Δ83hCG and $\alpha$ Δ 83FSH mutants had no secretion. These results are the first conclusive evidence that COOH-terminal amino acids are very important for secretion in human glycoprotein hormone $\alpha$-subunit. These results demonstrated that the $\alpha$ Δ83hCG and $\alpha$ Δ 83FSH mutants could be play a pivotal role in the secretion of tethered-molecule.

• 미생물학회지
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• 제27권2호
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• pp.77-84
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• 1989

Saccharomyces cerevisiae의 RNAI 유전자의 온도 감수성 돌연변이 균주는 성장허용 온도인 23"C에서는 정상적인 성"J--을하나, 성 장억제 온도인 36°C에서는 tRNA, rRNA 그러고 mRNA 의 선우울질을을 핵내에 축적함으후써 성장을 못한다. 본 실 험에서는 complementation 에 의하여 RNAI 유전자를 클로녕하였으며 concomitant loss 실험에 의하여 이 유천자의 클로닝 을 확인하였다. 유전자의 위치를 확인한 결과 3.5kb의 Bgl II 조각내에 RNAI 유전자가 존재함을 알 수 있였으며, 2.1kb에 해당하는 BamH I -Bgl II 조각에서는 RNAI 유전지에 의한 complementation 능력이 상실되는 것으후 보아 RNAI 유전자 는 3.5kb의 BglIl 조각내에 포함되는 BamH 1 자리 주위에 결쳐 있픔을 알 수 있었다.

• 한국정보통신학회:학술대회논문집
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• 한국해양정보통신학회 2002년도 춘계종합학술대회
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• pp.201-204
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• 2002

클라이언트에서 검색한 지도를 스크롤할 경우 이미 전송 딴은 객체를 캐슁하여 무선 데이터 전송량을 줄일 수 있지만, 클라이언트 내에서 데이터 좌표 변환, 선택적인 객체 삭제, 압축 단계가 부가적으로 요구된다. 특히 좌표 변환 연산은 모바일 휴대폰의 낮은 계산 능력, 적은 메모리 용량 등과 같은 제한적 자원으로 많은 시간이 요구된다. 따라서 휴대폰 기반 모바일 지도 서비스에서 효과적인 지도 제어 연산을 수행하기 위해서는 휴대폰의 제한적인 자원을 극복함과 동시에 무선 네트웍 대역폭을 줄이는 방안이 연구되어야 한다. 이 논문에서는 클라이언트-서버 시스템에서 이미 전송 딴은 객체의 효율적인 캐슁 기법을 제안하고자 한다. 또 캐슁된 데이터의 재이용과 원시 데이터의 재전송 방법의 장단점에 분석하고자 한다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제3권1호
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• pp.11-14
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• 1998

A toluene-oxidizing strain of Pseudomanas mendocina KR1 containing toluene-4-mono-oxygenase (TMO) completely degrades TCE with the addition of toluene as a co-substrate in aerobic condition. In order to construct in situ bioremediation system for TCE degradation without any growth-stimulating nutrients or toxic inducer such as toluene, we used the carbon-starvation promoter of Pseudomonas putida MK1 (Kim, Y. et al., J. bacteriol., 1995). Upon entry into the stationary phase due to the deprivation of nutrients, this promoter is strongly induced without further cell growth. The TMO gene cluster (4.5 kb) was spliced downstream of the carbon starvation promoter of Pseudomonas putida MK1, already cloned in pUC19. TMO under the carbon starvation promoter was not expressed in E. coli cells either in stationary phase or exponential phase. For TMO expression in Pseudomonas strains, tmo and carbon starvation promoter region were recloned into a modified broad-host range vector pMMB67HES which was made from pMMB67HE(8.9 kb) by deletion of tac promoter and lacIq (about 1.5 kb). Indigo was produced by TMO under the carbon starvation promoter in a Pseudomonas strain of post-exponential phase on M9 (0.2% glucose and 1mM indole) or LB. 18% of TCE was degraded in 14 hours after entering the stationary phase at the initial concentration of 6.6 ${\mu}$M in liquid phase.

• Journal of Microbiology and Biotechnology
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• 제10권5호
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• pp.690-693
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• 2000

Efficient intron deletion with the correct splicing of the two exons of the human proinsulin gene was accomplished by a novel stepwise method using genomic DNA [5]. The two exons were separately amplified in two steps, using the second step primers that incorporated additional bases complementary to the other exon. The fragments were combined in a third PCR reaction. Cloning and sequencing of the PCR product demonstrated the correct splicing of the two exons. Expression studies, using the pET9a vector, revealed a protein band with the correct size with respect to human proinsulin as confirmed by SDS-PAGe and Western blot. Proinsulin concentration was estimated to be around 200 mg per liter culture, expressed as inclusion bodies. Protein secretion to the culture medium and periplasmic space was achieved by cloning in the pEZZ18 vector.

• Journal of Microbiology
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• 제38권4호
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• pp.245-249
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• 2000

Catabolic pathways for the degradation of various aromatics by Sphingomonas yanoikuyae Bl are intertwined, joining at the level of substituted benzoates, which are further degraded vita ring cleavage reactions. The mutant strain EK497, which was constructed by deleting a large DNA region containing most of the genes for biphenyl, naphthalene, m-xylene, and m-toluate degradation, was unable to grow on all of the aromatics tested except for benzoate as the sole source of carbon and energy.S. yanoikuyae EK497 was found to possess only catechol ortho-ring cleavage activity due to deletion of the genes for the meta-cleavage pathway. Wild-type S. yanoikuyae Bl grown on benzoate has both catechol orthoand meta-cleavage activity. However, m-xylene and m-toluate, which are metabolized through methylbenzoate, and biphenyl, which is metabolized through benzoate, induce only the meta-cleavage pathway, suggesting the presence of a substrate-dependent induction mechanism.