• Korean journal of applied entomology
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• v.38 no.2
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• pp.145-149
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• 1999

To develop the baculovirus expression vector system (BEVS) adopting p10 gene promoter of Spodoptera exigua nucleopolyhedrovirus (SeNPV), we characterized the p10 gene of SeNPV. The nucleotide sequence of 545 bases including the coding region of p10 gene was determined. Compared with the previously reported SeNPV p10 gene (Zuidema et al., 1993), 4 bases were different in the 5' and 3' flanking region but no difference was found in the coding region. The p10 gene was located within Xho I 1.5 Kb, Sph 1 2.4 Kb and Cla I 4.0 Kb fragments by Southern hybridization analysis. Also, the Sph I 2.4 Kb and the Cla I 4.0 Kb fragments were cloned and their restriction enzyme maps were determined.

• Journal of Chest Surgery
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• v.34 no.1
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• pp.64-71
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• 2001

배경: 최근 치료법의 진보에도 불구하고 진행성 식도암의 예후는 5년 생존율이 10% 이하로매우 불량하기 때문에 식도암에 대한 새로운 치료방법의 하나로 암면역 치료가 대두되고 있다. 암면역 치료를 위해서 MAGE 등 종양 특이항원이 연구의 대상이 되고 있으나 국내에서는 아직 이에 대한 연구가 없다. 대상 및 방법: 1995년 1월부터 1998년 12월까지 고신대학교 복음병원 흉부외과에서 수술 치험한 125례의 식도암중 병리조직 보관상태가 양호한 편평세포암 79례를 병기에 따라(1병기 19례, IIa병기 19례, IIb병기 10례, III병기 21례, IV병기 10례) 무작위로 추출하고 대조군으로 평활근종 20례와 정상 식도점막 20례를 대조군으로 하여 DO7 단클론 항체와 항 MAGE-3 단클론 항체 57B를 이용하여 면역조직화학검사를 시행하여 변형 p53 단백과 MAGE-3 유전자 산물의 발현율을 조사하고 식도암 조직에서 질병의 진행도를 반영하는 병기에 따른 발현율 및 변형 p53 단백과 MAGE-3 유전자 산물의 발현율간의 상관관계를 조사하였다. 결과: 식도암조직에서 변형 p53 단백과 MAGE-3 유전자 산물의 발현율은 각각 51.9%, 60.8%의 발현율을 보였으나 식도평활근종과 정상 식도점막에서는 한례도 발현되지 않아 변형 p53 단백과 MAGE-3 유전자 산물은 대조군에 비해 식도암 조직에서 의미있게 발현되었다(p<0.001). 변형 p53 단백과 MAGE-3 유전자 산물의 발현은 I병기에서 68.4%, 52.6%, IIa병기에서 57.9%, 47.6%, IIb병기에서 60%, 70%, III병기에서 33.3%, 71.4%, IV병기에서 40%, 70% 각각 발현되어 병기에 따른 발현율의 차이는 없었다(p=0.193, p=0.452). 식도암 조직내에서 변형 p53 단백과 MAGE-3 유전자 산물의 발현간에는 상관관계가 없는 것으로 나타났다(p=0.679). 결론: 이상의 결과로 변형 p53 단백과 MAGE-3 유전자 산물의 발현은 식도암에서 예후인자로서의 역할은 할수 없으나 식도 편평세포 암조직에서만 특이하게 높은 빈도로 발현됨으로써 식도암도 면역치료의 대상이 될 수 있음을 확인하였다.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.16-16
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• 2001

Apoptosis로 일컬어지는 예정된 세포사멸(programmed cell death)은 개별 세포의 입장에서는 곧바로 사멸을 의미하지만, 정상적인 고등 생물의 입장에서는 개체의 발생과 분화하는데 프로그램된 과정이다. 자발적 세포사멸은 다른 조직에 비해 생식 조직인 난소나 정소에서 복잡한 apoptosis 기작들을 가지리라 사료된다. 본 연구는 Bcl-2 family중 apoptotic protein인 Bax에 대해 suppression하는 유전자를 yeast system을 활용하여 돼지 정소와 난소로부터 각각 cDNA library를 구축한 후 탐색하였다. 탐색에 활용된 cDNA library는 돼지의 정소와 난소로부터 mRNA를 분리하여 yeast vector인 pAD-GAL4-2.1에 구축하였고, 마우스 bax 유전자는 gal 1 promoter의 조절 하에 glucose 배지에서는 유도되지 않고, galactose 배지에서만 선택적으로 Bax를 발현할 수 있는 효모 vector(pL19-bax)를 구축하였다. Bax에 의한 apoptosis suppressor를 탐색하기 위해 우선 효모 W303에 pL19-bax를 transform하여 glucose 배지에서 Bax의 발현을 억제하였다. pL19-bax를 가진 효모에 정소와 난소로부터 구축된 cDNA library를 transform 시키고, transform된 효모는 각각 Bax에 의한 toxicity를 저해하는 유전자를 찾기 위해 스크린되었다. 이러한 방법으로 정소 cDNA library 탐색에서는 5 $\times$ $10^{6}$ transformant중 39개, 난소cDNA library 탐색에서는 2 $\times$ $10^{6}$ transformant중 26개의 콜로니가 생존하였다. 이들 콜로니로부터 유전자를 분리하여 분석해 본 결과 여러 그룹으로 분류할 수 있었다. 각 그룹의 관련 유전자는 protein synthesis/degradation 12종, oxidation/reductation 5종, detoxin/ cell cycle promoter 3종, signal transduction/growth factor 5종, 그리고 알려지지 않은 유전자 9종이었다. 그 중, bax-toxicity inhibition에 강력한 survival phenotype을 가지는 유전자(pSEDL)를 동정하였다. 이것은 T3-4-1 콜로니로부터 분리하였는데 140개 아미노산으로 이루어진 인간 SEDL(GenBank, XM_013096) 유전자와 매우 유사한 homology를 가지며, bax와 관련된 기능은 밝혀져 있지 않다. 이외에도 분리된 유전자에는 NADH, thioreduction, 그리고 cytochrome oxidase와 같은 positive 유전자 군이 크로닝되어, Bax를 이용한 효모에서 apoptosis suppressor에 관련된 유전자를 손쉽게 스크린하는 것이 가능하고, 분리된 유전자의 기능을 예측할 수 있어 지금까지 보고된 유전자 크로닝법 보다는 강력한 수단으로 활용될 수 있다는 사실을 시사하였다. 그러나, ORF에 관계없이 Bax 발현에 저항하는 유전자군이 선발된다든지 하는 문제점은 금후 검토가 필요하리라 사료된다.

• Korean Journal of Microbiology
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• v.52 no.3
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• pp.298-305
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• 2016

Previously our laboratory showed that Pseudomonas alkylphenolica KL28 possesses two different lap and pcu gene clusters for p-cresol catabolism. In this study, additional gene cluster (pchACXF-pcaHG-orf4-pcaBC) has been identified to encode enzymes necessary for catabolism of p-cresol to ${\beta}$-carboxy-cis,cis-muconate. This gene cluster showed almost identical nucleotide sequence homologies to those in the plasmid of Pseudomonas putida NCIMB 9866 and 9869, British origins, indicating the possibility of a horizontal gene transfer. Through mutagenesis of each gene cluster and gfp-based promoter reporter assays, it has been shown that the three gene clusters are functionally operated and pch genes are induced by p-cresol. Furthermore, the pcu gene cluster of the three was shown to be dominantly expressed in utilization of p-cresol. Mutation of the pcu gene was defective in aerial structure formation under p-cresol vapor, indicating the utilization rate of carbon source is one of key elements for the multicellular development of this strain.

• The Journal of the Korea Contents Association
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• v.17 no.10
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• pp.289-299
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• 2017

To verify the association between susceptibility to chronic myeloid leukemia (CML) and GSTM1, GSTT1 gene polymorphisms in Asian populations, 9 papers published until July 2017 were cited in a meta-analysis. The null present types of the GSTM1, GSTT1 gene were analyzed individually. The significant association was found between CML and GST polymorphism (GSTM1; OR=1.306, 95% CI=1.091-1.563, p=0.004, GSTT1; OR=1.987, 95% CI=1.438-2.746, p=0.000). In addition, there was association between CML and the null type of the combination GSTM1-GSTT1 polymorphisms (OR=4.191, 95% CI=2.833-6.201, p=0.000). Thus, genetic polymorphisms of the GSTM1, GSTT1 and combination GSTM1-GSTT1 polymorphism in Asian populations may be risk factors for CML.

• Journal of Aquaculture
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• v.10 no.1
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• pp.33-37
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• 1997

construct containing reporter gene(pFV4CAT) regulated by carp $\beta$-actin promoter was microinjected into the one-cell stage egg of mud loach (Misgurnus mizolepis), and was successfully expressed, possibly by the integration into the genome. Both mean hatching success and early survival of the microinjected groups were not significantly different with those of control groups (P>0.05). The incidence of transgene was ranged from 7 to 48% based on the PCR and/or Southern blot analyses with the DNA prepared from fin or blood tissue. The spatial and temporal patterns of expression of the pFV4CAT gene, measured by in situ immunohistochemical analysis peroxidase-conjugated anti-CAT antibody, were variable among the experimental individuals. These results suggest that carp $\beta$-actin promoter is effective to express other transgene in mud loach, such that this promoter can be useful in the generation of valuable transgenic mud loach.

• Journal of Plant Biotechnology
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• v.34 no.4
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• pp.307-312
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• 2007

Chlamydomonas reinhardtii was transformed with the coding sequence of the Tombus virus gene p19 to determine whether the gene functions as an RNAi suppressor in C. reinhardtii. Transformants were confirmed to have 1 to several copies of p19 gene in their chromosomes. When an RNAi strain of C. reinhardtii generated by transforming the inverted repeat (IR) sequence homologous to the 3'UTR region of the MAA7 gene was re-transformed with the gene p19, MAA7 transcript levels of transformants fluctuated and proliferation of trans-formants on the medium containing 5-FI was suppressed. Overall results suggest that p19-mediated silencing suppression works at a low level in C. reinhardtii because of difference in codon usage resulting in weak P19 expression unless p19-mediated silencing suppression in C. reinhardtii works in a different manner from higher plants.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.266-266
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• 1994

$B^{30}$ 위치에 homoserine이 치환된 사람 insulin 유사체 ($B^{30}$ -homoserine) insulin을 생산하기 위해, insulin의 B 사슬 유전자에 A 사슬 유전자를 직접 연결한 insulin 유전자를 설계하였다. 이 유전자는 10개의 oligonucleotide로 나누어 합성하여 T4 DNA ligase로 결합시킨 후, pUC19 plasmi의 polylinker 영역에 삽입하였다. 이 유전자의 발현을 높이기 위해 이 유전자는 다시 tac promoter의 지배를 받는 lacZ 유전자의 Cia I 또는 Hpa I 제한부위에 도입하여 융합시켰다. 이렇게 구축된 운반체 pTBA나 pKBA를 Escherichia coli JM103 균주에 형질도입시킨 후, 이를 4시간 배양한 후 0.05mM이상의 isopropyl-$\beta$-D-thiogalactopyranoside (IPTG)를 배지에 공급해 주고 2시간 더 배양하였을 때 유전자 발현이 잘 유도되어짐을 알 수 있었다. 이 때 생산된 insulin 전구체들은 세포내 불온성인 inclusion body로 축적되어지는 것을 관찰하였으며, 그 생산량은 세포내 전체 단백질량의 30%에 달하였다.

• Journal of Life Science
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• v.28 no.11
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• pp.1277-1284
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• 2018

In this study, a repeated yeast integrative plasmid (R-YIp) harboring Cre/loxP system was constructed to integrate various gene expression cassettes into the yeast chromosome. The R-YIp system contains a reusable selective marker (CgTRP1), loxP sequence, and target sequence for integration. Therefore, many gene expression cassettes can be integrated into the same position of the same yeast chromosome. In the present study, several model enzymes involving xylan/xylose metabolism were examined, including endoxylanase (XYLP), ${\beta}$-xylosidase (XYLB), xylose reductase (GRE3) and xylitol dehydrogenase (XYL2). Efficient expression of these genes was obtained using two promoters (GAL10p and ADH1p) and various plasmids (pGMF-GENE and pAMF-GENE plasmids) were constructed. The XYLP, XYLB, GRE3, and XYL2 genes were efficiently expressed under the control of the GAL10 promoter. Subsequently, R-YIps containing the GAL10p-GENE-GAL7t cassette were constructed, resulting in pRS-XylP, pRS-XylB, pRS-Gre3, and pRS-Xyl2 plasmids. These plasmids were sequentially integrated into chromosome VII of a Saccharomyces cerevisiae strain by repeated gene integration and selective marker rescue. These genes were integrated by the R-YIp system and were stably expressed in the yeast transformants to produce active recombinant enzymes. Therefore, we expect that the R-YIp system will be able to overcome current limitations of the host cells and allow selective marker selection for the integration of various genes into the yeast chromosome.

• Korean Journal of Microbiology
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• v.30 no.5
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• pp.371-376
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• 1992

Plasmid pSL4 of plasmid pKM 101 mutant have high protection effects and mutagenecity for UV and methyl methanesulfonate, The mucA gene and a pan of mucE gene of pKM 101 and pSL4 were sucloned onto lacZ' fusion vector pMC874 and the hybrid plasmids pBH31 and pBH30 were selected. These plsmids were intrduced into $recA^{+}lexA^{-}$, $recA^{-}와lexA^{+}$ strains and determined the activity of $\beta$-galactosidase for UV. In $recA^{+}lexA^{+}$ strain.$\beta$-galactosidase activity of pBH30 included mue region of pSL4 was higher thall pBH31 inclued muc region of pKM 10 I and the tf-galactosidase of two plasmids was not induced in reeA and leeA mutants with or without UV illumination. Without UV illumination. the .$\beta$-galactosidasc of pBH30 was expressed a little higher level than that of pBH3L We suggest that the functional difference of pKM 10l and pSL4 are due to the variety of mue regulatory region. Also. a plasmid pBH 100 earring umuC' -lacZ' gene fusion was constructed in vitro to study the regulation of the umu operon. It was shown that the umu operon is induced by UV and is regulated by the reeA and lexA genes.