Yeast Artificial Chromosome의 효율적인 조작과 분석을 위한 새로운 Bicistronic Fragmentation Vector의 개발에 관한 연구

임향숙,최주연,김인경,강성만,성영모 한국미생물학회 1999 미생물학회지 Vol.35 No.1

Fragmentation vectors are used to analyze function and genomic structure of a gene of interest by creating deletion derivatives of large fragments of genomic DNA cloned as yeast artificial chromosomes (YACs). Herein, we developed a new hicistronic fragmentation vector that contains internal ribosomal entry sile (IRES) of encephalomyocarditis vin~s (EMCV) and $\beta$-galactosidase as a reporter gene. This vector system provides a novcl loo1 to analyze expression patterns of a gene of interest due to simultaneous expression of a target gene as well as $\beta$-galactosidase driven from a single message. In addition, the bicistronic fragmentation vector contains four rare-cutting restriction enzyme sites in the polycloning sites which can be used to conveniently insert any kinds of genes and therefore facilitates targeting DNA scgments into YAC by means of homologous recombination. This approach establishes a paradigm for manipulation of mammalian DNA segments and characterization of expression and regulatory regions of mammalian gene cloned as YAC. 본 연구에서는 Yeast Artificial Chromosome을 효율적으로 조작하고 유유전자의 기능을 보다 더 용이하게 분석하기 위해 EMCV 바이러스의 IRES 염기서열과 $\beta$-galactosidase를 포함하는 새로운 bicistronic fragmentation vector를 제조하였다. 이 벡터의 폴리클로닝 site에 네 개의희귀한 제한효소 부위를 도입하여 DNA를 용이하게 클로닝할 수 있게 만들었다. 그러므로 원하는 어떤 YAC도 효모세포에서 쉽게 상동 재조함에 의해 절편할 수 있는 장점이 있다. 이 bicistronic fragmentation vector 시스템을 이용하면하나의 메시지로부터 연구하고자 하는 유전자와 $\beta$-galactosidase를 동시에 한 세포에서 발현할 수 있기 때문에 유전자의 발현양상을 쉽게 분석할 수 있는 새로운 도구로 이용할 수 있다.

지역사회복지협의회의 복지사각지대 해소를 위한 좋은이웃들

임향숙 한국지역사회복지학회 2016 한국지역사회복지학회 학술대회 Vol.2016 No.05

Methylase를 사용한 Escherichia coli에서 Triplex 존재에 관한 연구

임향숙,김성조,강성만,Rhim, Hyangshuk,Kim, Sungjo,Kang, Seongman 한국미생물학회 1998 미생물학회지 Vol.34 No.4

Escherichia coli 세포에서 intramolecular triplex가 형성될 수 있는지를 조사하기 위하여 genetic assay를 도입하였다. Two dimensional gel electrophoresis 방법을 사용하여 $(G)_9AATTC(G)_9$ 혹은 $(GAA)_9TTC(GAA)_8$ sequences가 in vitro에서 intramolecular triplex 구조를 형성하는 것을 확인하였다. Genetic assay를 위하여 temperature-sensitive EcoRI methylase 유전자를 포함한 플라스미드와 triplex를 형성할 수 있는 $(G)_9AATTC(G)_9$ 혹은 $(GAA)_9TTC(GAA)_8$ sequences를 포함한 플라스미드로 E. coli를 cotransform 하였다. EcoRI methylase가 발현되는 permissive temperature에서 pur pyr sequences를 포함하는 플라스미드는 EcoRI methylase 작용에 kinetically 저항성을 보여주었다. 이러한 결과는 pur pyr sequence들이 EcoRI methylase가 작용하기 어려운 triplex 구조를 E. coli 세포에서 형성한다는 것을 증거한다. We have introduced a genetic assay to study the existence of intramolecular triplexes in Escherichia coli. A plasmid containing the gene that encodes a temperature-sensitive EcoRI methylase was cotransformed with different plasmids containing inserts, $(G)_9AATTC(G)_9$ and $(GAA)_9TTC(GAA)_8$, that are able to form intramolecular triplexes in vitro. Inhibition of methylation in vivo was found for $(G)_9AATTC(G)_9$ and $(GAA)_9TTC(GAA)_8$, suggesting that the pur pyr sequences adopt unusual strucures in E. coli. In addition, experiments using two dimensional gel electrophoresis confirmed that intramolecular triplexes are formed for the pur pyr sequences under negative supercoiling. These results demonstrate the existence of intramolecular triplexes in E. coli.

임상간호사의 전문직관과 조직몰입과의 관계

임향숙,오경옥 충남대학교 간호과학연구소 1999 충남대 간호학술지 Vol.2 No.1

The purpose of this study was to identify the relationship between views on profession and organizational commitment. The sample for the study consisted of 268 nurses of two University Hospitals in Taejon & Cheongju. Data collection period was from March 3,1998 to March 11,1998. The tools used for this study were composed of General characteristics^ items), Views on profession(33 items), Organizational commitment (15 items). The data was analyzed by using an SAS program and included frequency, percentage, mean, t-test, ANOVA, Pearson correlation coefficient, Multiple regression. the finding were as follows: 1. The mean score of views on profession was 3.357, organizational commitment was 2.284 on a 5 point Likert scale. 2. General characteristics were showed significant difference in views on profession: age(F=3.37 p=.013), position(F=4.34 p=.014), marital status(t=3.58 p=.000). 3. General characteristics were showed significant difference in organizational commitmnt: age(F=9.87 p=.000), education level(F=7.00 p=.ooo), position(F=20.52 p=.000), working experience(F=8.68 P=.000), marital status (t=3.86 p=.000), religion(F=2.50 p=.043). 4. General characteristics were statistically significantly correlated with views on profession:age(r=0.165 p=.006), position(r=0.172 p=.004), marital status(r=0.214 p=.000). 5. General characteristics were statistically significantly correlated with organizational commitment: age ( r = 0.236 p = .000), position (r = 0.359 p = .000), working experience(r=0.218 p=.000), marital status(r=0.230 p=.000). 6. There was a positive relationship between views on profession and organizational commitment(r=.554 p=.000). Based on the study results, the following recommendation are made: 1) Future research will be promoted valid and reliable tool development. 2) A future study is necessary to develop other variable which positively influence.

CHEMICAL SYNTHESIS AND CLONING OF HUMAN β - ENDORPHIN GENE

이현주,임향숙,이대실,김명희 한국생화학분자생물학회 (구 한국생화학회) 1987 추계학술대회집 Vol.- No.-

Study of Human Insulin : I. Synthesis and Cloning of Genes for Human Insulin A and Chains

박병철,임향숙,김명희,이대실,Park, Byung-Chul,Lim, Hyang-Sook,Kim, Myoung-Hee,Lee, Dae-Sil 생화학분자생물학회 1988 한국생화학회지 Vol.21 No.3

인슈린을 박테리아에서 발현시키기 위해 인체 인슐린 A와 B 사슬에 해당하는 유전자를 설계하였다. 화학적 합성, 생화학적 조립, 유전자 재조합상 용이성 등이 고려된 설계에는 각 인슈린 사슬이 11개의 유전자 절편으로 구성되도록 하였다. 계획된 유전자 절편은 phosphite triester 합성법으로 합성하였으며, 인슐린의 A와 B 사슬의 합성유전자를 건설하기 위해서 합성된 유전자 절편을 효소적인 방법으로 결합하였고, 이어서 대장균에 클로닝 하였다. 클론들은 계획한대로 인슐린 유전자가 대장균에 도입되었음을 유전자 서열 결정에 의해 확인하였다. Total synthesis of human insulin A and B chain genes were designed for the expression through bacterial system. Eleven individual oligonucleotides corresponding to insulin A and B chains were chemically synthesized by the phosphite triester method on solid support. The enzymatic joining of the oligonucleotides were employed to give the synthetic human insulin A and B chain genes, which ware separately cloned into the bacterium Escherichia coli using plasmid, pUC19. The cloned DNAs were shown to have the correct nucleotide sequence as designed.

인체 인슈린 연구 . 1 . 인체 인슈린 A 와 B 사슬의 유전자합성과 클로닝

박병철,임향숙,김명희,이대실 ( Byung Chul Park,Hyang Sook Lim,Myoung Hee Kim,Dae - Sil Lee ) 생화학분자생물학회 1988 BMB Reports Vol.21 No.3

Total synthesis of human insulin A and B chain genes were designed for the expression through bacterial system. Eleven individual oligonucleotides corresponding to insulin A and B chains were chemically synthesized by the phosphite triester method on solid support. The enzymatic joining of the oligonucleotides were employed to give the synthetic human insulin A and B chain genes, which ware separately cloned into the bacterium Escherichia coli using plasmid, pUC19. The cloned DNAs were shown to have the correct nucleotide sequence as designed.

제10차 춘계 학술대회 초록집 : 포스터발표 1 ; 간암세포에서 prostaglandin (PG) A2와 △(12)-PGJ2에 의해 유도된 Sox4 유전자의 기능 연구

허원희,임향숙,왕진상,배시현,최종영,양진모,왕희정,윤승규 대한간학회 2004 Clinical and Molecular Hepatology(대한간학회지) Vol.10 No.3(S)

자기공명분광법에서 TE와 Voxel 내의 대사물질 양에 따른 스펙트럼 변화 평가에 관한 연구

우동철,김상수,임향숙,장건호,최보영,Woo, Dong-Cheol,Kim, Sang-Soo,Rhim, Hyang-Shuk,Jahng, Geon-Ho,Choe, Bo-Young 한국의학물리학회 2007 의학물리 Vol.18 No.3

본 연구는 자기공명분광법(Magnetic Resonance Spectroscopy: MRS)에 있어서 기존 개발된 Multi-Voxel Spectroscopy (MVS)을 위한 MR 원뿔형 팬톰을 Single voxel spectroscopy (SVS)용으로 개선하여 선정된 Voxel내의 대사물질의 양과 TE 변화에 따른 스펙트럼의 변화를 관찰하였다. 원뿔형 팬톰은 Voxel를 선정하는 위치에 따라서 그 안에 포함되는 대사물질의 양을 변화시킬 수 있다는 점에 착안하여 Voxel안에 포함되는 뇌대사물질의 양과 TE시간에 따른 각 대사물질(NAA, Cr, Cho, Lac 등)의 Peak에 대한 정량적인 분석을 시도하였다. 실험은 3T MRI/MRS 장비에서 이루어졌고, 데이터를 분석하는데 $jMRUI^{(R)}$라는 프로그램이 사용되었다. 실험 및 분석 결과 대체로 Echo Time (TE)가 커질수록 잡음이 감소하는 경향을 확인할 수 있었고 또한 TE가 커질수록, Voxel 안의 뇌대사물질의 양이 적을수록 각 대사물질의 peak intensity가 감소하는 것을 관찰할 수 있었다. 특히 Lactate의 경우, 정위선정된 Voxel안에 가장 많은 양의 대사물질이 포함되었을 때만 스펙트럼 분석이 가능한 정도의 Peak intensity를 얻을 수 있었고 대부분의 TE에서 분석 가능한 것으로 보아 각 대사물질의 intensity는 TE보다는 Voxel내의 대사물질의 양에 더욱 민감하다는 결론을 도출 할 수 있었고, 이러한 Lac에 대한 in vitro 데이터는 정량분석 시에 있어서 in vivo상에서 대사물질의 정량화를 하는 데 있어서 중요한 가이드라인을 제시할 것으로 예상된다. 그러나, 원뿔의 모서리 부분에 표면장력에 의하여 수많은 공기방울이 MR image상에서 관찰되고 또한 MRS상에서 그 스펙트럼을 얻을 수 없었는데 이는 앞으로 원뿔형 MRS 팬톰을 구조적, 기술적으로 더욱 개선해야 할 여지를 남겨두고 있다. The purpose of this study is to investigate the spectra of a magnetic resonance spectroscopy (MRS) in accordance with the variance of TE and the volumes of metabolites in a localized voxel for the quality assurance using a designed single voxel spectroscopy QA phantom. Because a cone-shade phantom is designed as the volume of metabolite in a localized voxel is changeable, we try to analyze the peaks of each metabolite (NAA, Cr, Cho, Lac, etc.) in accordance with metabolite volume in a localized voxel as well as echo time (TE). All data were obtained using a 3T MRI/MRS machine and analyzed using $jMRUI^{(R)}$. The results of this study show that TE is in inverse proportion to the noise of MRS and the longer TE and the less metabolite volume in the localized voxel, the peak intensities of each metabolite decrease. In case of the lactate, its peak was observed on the all TE only if the greatest metabolite is included in the localized voxel. Then, the intensity of a metabolite is more sensitive to the metabolite volume in the localized voxel than the TE. These obtained in vitro MRS data is provide the guideline that is important for in vivo metabolite quantification. But, in the edge of cone-shape vial air bubbles were observed and spectrum could not obtained. Therefore our cone-shape MRS phantom needs to be modified in order to solve these problems.

Hip2 ubiquitin-conjugating enzyme has a role in UV-induced G1/S arrest and re-entry

홍난희,탁영진,임향숙,강성만 한국유전학회 2019 Genes & Genomics Vol.41 No.2

Regulation of cell cycle arrest and re-entry triggered by DNA damage is vital for cell division and growth and is also involved in cell survival. UV radiation can generate lesions in the DNA, which results in cell cycle arrest and the induction of the DNA repair process. However, the mechanism of promoting cell cycle progression following DNA repair is elusive. The primary aim of this study is to investigate whether Hip2 ubiquitin-conjugating enzyme has a role in UV-induced G1/S arrest and re-entry. The phase of HEK293 cells was synchronized at the G1/S border using thymidine. The synchronously proliferating cells were exposed to UV radiation to cause DNA damage. We investigated the expression of p53, Hip2, p21, cyclin D and E proteins that are involved in the cell cycle progression. Finally, we examined changes in the phosphorylation of Hip2 after UV radiation treatment using the pIMAGO™ assay. When cells were exposed to UV radiation, expression of p53 was elevated, and the cell cycle was arrested at the G1/S boundary. In response to the increased p53 level, Hip2 became phosphorylated and activated through the inhibition of its degradation. The phosphorylated Hip2 inhibited p53, thereby suppressing the expression of p21, a downstream signal, and sequentially stimulating cyclin D and cyclin E to induce reentry to the cell cycle. Our studies demonstrate that Hip2 works as a regulator in UV-induced cell cycle arrest and re-entry.