• 생명과학회지
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• 제6권3호
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• pp.185-192
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• 1996

박테리오파지 T7 gene 2.5 단백질은 single-stranded DNA 결합 단백질로 박태리오파지 T7의 DNA복제, 재조합, 및 수선에 필수적으로 요구된다. Gene 2.5 protein은 T7의 DNA 합성과 성장에 필수적인 단백질이다. Gene 2.5 Protein이 중요시 되는 이유는 이 단백질이 T7의 다른 복제 필수단백질인 T7의 다른 복제 필수단백질인 T7 DNA polymerase 와 gene 4 protein(helicase/primase)와 서로 상호작용할 것으로 제안되었기 때문이다. (Kim and Richardson, J. Biol. Chem., 1992;1994). 이 단백질의 단백질 상호작용을 가능하게 하는 domain은 carboxyl-terminal domain일 것으로 여러 실험에서 대두되었기에, 이 domain의 특성을 파악하기 위해 야생형과 변이체 gene 2.5 단백질들을 각각 GST에 융합한후 fusion 단백질을 정제하였다. 정제된 이 융합 단백질들의 carboxyl-terminal domain이 T7 복제 단백질들과 상호작용을 조사하는지를 조사하기 위해 affinity chromatography로 이용하였다. 실험 결과, 아생형 GST-gene 2.5 융합단잭질(GST-2.5 (WT))는 T7 DNA polymerase 와 상호작용을 하였지만. 변이형 융합단백질(GST-2.5$\Delta$21C)는 interaction을 하지 못했다. 이 결과는 carbohyl-terminal domain이 단백질-단백질 상호작용을 하는데 직접적으로 관여하는 것을 증명하였다. 또한,GST2.5(WT)는 gene 4 protein(helicase/primase)와 직접 상호작용을 하나. GST2.5$\Delta$21C는 상호작용을 하지 못하는 것으로 나타났다. 따라서 gene 4 proteins와의 상호작용에도 gene 2.5 protein의 carboxyl-terminal domain이 직접 관여 한다는 것이 증명되었다. 이상의 결과에서 gene 2.5 protein은 박테리오파지 T7 의 유전자 목제 시 단백질-단백질 상호작용에 관혀아며, 특히 gene 2.5 protein의 carboxyl-terminal domain이 이러한 상호작용에 직접적으로 관여하는 domain이라는 것을 알 수가 있었다.

• 한국가축번식학회지
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• 제27권3호
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• pp.225-231
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• 2003

본 연구는 생체 내 세포 및 조직배양기로서의 면역결핍동물을 개발할 목적으로 proximal lck promoter와 DT-A gene를 이용하여 형질전환생쥐을 생산하고 이 형질전환생쥐의 면역세포에서 DT-A gene이 발현되는지를 분석하였다. 형질전환생쥐와 정상생쥐로부터 thymus, spleen 및 liver에서 RNA를 추출하여 RT-PCR수행하였는데 정상생쥐의 조직에서는 어떠한 DT-A gene의 발현양상을 얻을 수 없었으나 형질전환생쥐의 thymus, spleen, liver에 DT gene의 발현을 확인할 수 있었고, Northern blotting을 이용하여 형질전환생쥐의 thymus, spleen 및 liver에서 DT-A gene이 강하게 발현되는 것으로 나타났다. 형질전환생쥐 $F_1$ 및 $F_2$ 산자의 혈액에서 T-cell 발달의 분포도를 확인하기 위해 CD4 및 CD8 antibody를 이용하여 FACS analysis를 실시하였는데 형질전환생쥐의 혈액 내 mature T-cell인 single positive thymocyte의 수가 정상생쥐에 비해 감소하는 경향을 나타냈다. 정상생쥐의 혈액 내 T-cell 중 $CD8^{+}$ T-cell의 경우 약 50%를 나타냈으나 형질전환생쥐의 경우 33%로 감소하였고, $CD4^{+}$ T-cell은 정상생쥐에서 10%를 차지하고 있으나 형질전환생쥐에서는 5.9%로 감소되는 것으로 분석되었다. 그러므로 본 연구의 형질전환생쥐에서 lck promoter에 의해 초기 immature한 상태의 T-cell에서 DT-A gene이 발현되어 발육중인 T-cell이 파괴 되어 mature 상태인 $CD4^{+}CD8^{-}$나 $CD4^{-}CD8^{+}$ cells (single-positive)들이 감소된 것으로 확인되었다.

• Journal of the Korean Data and Information Science Society
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• 제23권3호
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• pp.467-474
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• 2012

인간의 질병과 가축의 경제적인 특성은 하나의 유전자가 아닌 여러 유전자의 상호작용의 영향을 더 많이 받는다고 알려져 있다. 본 논문에서는 유전적인 효과만을 밝혀내기 위해 선형회귀모형을 활용하여 환경적인 요인을 보정하고, 최근 한우의 맛과 육질에 영향을 준다고 밝혀진 단일염기다형성 5개 (Oh 등, 2011)를 이용해 한우의 경제형질에 영향을 주는 우수 유전자 조합을 선별하고 우수 유전자형을 밝힌다. 이때, 많은 유전자들 중에서 우수한 유전자를 찾기 위한 비모수적인 방법인 다중인자 차원 축소 방법을 이용하여 단일 유전자의 효과보다 상호작용의 효과가 한우의 경제형질에 더 많은 영향을 준다는 사실을 확인하였다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권3호
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• pp.781-784
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• 2012

The aim of this study was to evaluate the effect of the adenovirus-mediated double suicide gene (CD/TK) for selective killing of gastric cancer cells. Gastric cancer cells SCG7901 and normal gastric epithelial cell lines were infected by adenoviruses Ad-survivin/GFP and Ad-survivin/CD/TK. GFP expression and CD-TK were detected by fluorescence microscopy and reverse transcriptase polymerase chain reaction (RT-PCR), respectively. After treatment of the infected cells with the pro-drugs ganciclovir (GCV) and/or 5-FC, the cell growth status was evaluated by methyl thiazolyl tetrazolium assay. Cell cycle changes were detected using flow cytometry. In nude mice bearing human gastric cancer, the recombinant adenovirus vector was injected directly into the tumor followed by an intraperitoneal injection of GCV and/or 5-FC. The subsequent tumor growth was then observed. The GFP gene driven by survivin could be expressed within the gastric cancer line SCG7901, but not in normal gastric epithelial cells. RT-PCR demonstrated the presence of the CD/TK gene product in the infected SCG7901 cells, but not in the infected normal gastric epithelial cells. The infected gastric cancer SCG7901, but not the gastric cells, was highly sensitive to the pro-drugs. The CD/TK fusion gene system showed significantly greater efficiency than either of the single suicide genes in killing the target cells (P<0.01). Treatment of the infected cells with the pro-drugs resulted in increased cell percentage in G0-Gl phase and decreased percentage in S phase. In nude mice bearing SCG7901 cells, treatment with the double suicide gene system significantly inhibited tumor growth, showing much stronger effects than either of the single suicide genes (P<0.01). The adenovirus-mediated CD/TK double suicide gene driven by survivin promoter combined with GCV an 5-FC treatment could be an effective therapy against experimental gastric cancer with much greater efficacy than the single suicide gene CD/TK combined with GCV or 5-FC.

• Journal of Plant Biotechnology
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• 제8권1호
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• pp.9-14
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• 2006

As an alternative method to produce low cost reagents for immunodiagnosis and protect the plants from viral disease, a gene encoding a single chain variable fragment(scFv) recombinant antibody targeted to the coat protein of cucumber mosaic virus (CMV) was expressed in Nacotiana tabacum. The source of the scFv recombinant antibody gene was from spleen tissue of an immunized mouse. The gene was initially cloned into the pCANTAB5E phagemid and expressed in E. coli. In the following study, the antibody gene was subcloned into the plant expression vector, pCAMBIA-1301 and introduced into tobacco leaf tissue via Agrobacterium tumefacients mediated transformation. After transformation, 56 out of 58 plants were shown to carry the desired anti-CMV scFv gene by PCR analysis. Overall, only 12.5% of the 56 putative transgenic plants were found to express the antibody to a detectable level.

• Journal of Microbiology and Biotechnology
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• 제18권2호
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• pp.334-337
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• 2008

A tightly regulated gene expression system composed of a single-copy target gene under the control of a lac promoter derivative and lacI gene in a multicopy plasmid is proposed, and its ability to control the flux of a metabolic pathway is demonstrated. A model system to control the flux of acetyl-CoA to acetyl phosphate was constructed by integrating pta, a gene encoding phosphotransacetylase, under a tac promoter into the chromosome of E. coli with a pta-negative background and transforming a multicopy plasmid containing the $lacI^Q$ gene into the strain. The production rate of acetate was shown to be tightly controlled when varying the concentration of the inducer (IPTG) in he model system.

• Journal of Genetic Medicine
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• 제3권1호
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• pp.15-19
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• 1999

We have investigated the genetic variation in the human apo B mRNA editing protein (apobec-1) gene. Exon 3 of the apobec-1 gene was amplified by polymerase chain reaction. After detection of an additional band by single strand conformational polymorphism (SSCP) analysis, sequencing of the SSCP-shift sample revealed a single-base mutation. The mutation was a CGG transversion at codon 80 resulting in a lleRMet substitution. This substitution was confirmed by restriction fragment length polymorphism analysis since a Pvull site is abolished by the substitution. Population and family studies confirmed that the inheritance of the genotypes for apobec-1 gene polymorphism is controlled by two codominant alleles (P1 and P2). A significant difference in plasma triglyceride was detected among the different apobec-1 genotypes in the CAD patients (P<0.05). Our study could provide the basis for elucidating the interaction between genetic variation of the apobec-1 gene and disorders related to lipid metabolism.

• 한국육종학회지
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• 제42권1호
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• pp.23-27
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• 2010

The objective of this study was to map the gene for reduced embryo size in rice using DNA markers. The reduced embryo size mutant was induced from N-methyl-N-nitrosourea (MNU) treated Taichung 65. Genetic analysis revealed that the phenotype of the reduced embryo was controlled by a single recessive gene, designated as re-1. For mapping the gene controlling embryo size, an $F_2$ population was developed from a cross between the Korean Tongil-type, Milyang 23 (Oryza sativa ssp. indica) and the mutant. The ratio of $F_2$ seeds nearly fitted to 3:1 ratio, indicating that this phenotype was controlled by a single recessive gene. Bulked sergeant analysis was performed with SSR markers. The gene for the reduced embryo size was detected on chromosome 1. The gene was further mapped between two SSR markers, RM315 and RM265 on chromosome 1 (approximately 1.5 Mb interval). The linked markers will facilitate selection of this grain character in a breeding program and provide the foundation for positional cloning of this gene.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2005년도 창립 30주년 및 춘계학술대회
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• pp.18-18
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• 2005

• Biotechnology and Bioprocess Engineering:BBE
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• 제6권2호
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• pp.117-127
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• 2001

The high-level expression of a human single chain urokinase-type plasminogen activator (scu-PA) was achieved by employing a methotrexate (MTX)-dependent gene amplification system in Chinese hamster ovary (CHO) cells. By cotransfecting and coamplifying a scu-PA expression plasmid and dihydrofolate reductase (DHFR) minigene, several scu-PA expressing CHO cell lines were selected and gene-amplified. These recombinant cell lines, NGpUKs, secreted a completely processed scu-PA of 54 kD and up to 60mg/L was accumulated in the culture medium when they were adapted to an optimal MTX concentration. Over 95% of the scu-PA expressed was secreted in the culture medium and identified having the proper function of a plasminogen activator when activated by plasmin. Based on a genomic Southern analysis, a representative subclone, MGpUK-5, exhibited MTX-dependent scu-PA gene amplification, plus the initial single-copy gene of scu-PA eventually turned into about 150 copies of the amplified gene of scu-PA after gradual adaptation to 2.0$\mu$M of MTX. Meanwhile, the transcripts kof the scu-PA gene increased, although -early saturation of transcription was identified at 0.1$\mu$M of MTX. The scu-PA production by the MGpUK-5 subclone also increased relative to the gene amplification and increased transcripts, however, the relationship was not linearly proportional. Accordingly, since the MGpUK cell lines expressed elevated levels of enzymatically active scu-PA, these cell lines could be applied to the largescale production of scu-PA.