• BMB Reports
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• 제35권6호
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• pp.623-628
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• 2002

The Th1 vs. Th2 balance is critical for the maintenance of immune homeostasis. Therefore, the genes that are selectively-regulated by the Th1 and Th2 cytokines are likely to play an important role in the Th1 and Th2 immune responses. In order to search for and identify the novel target genes that are differentially regulated by the Th1/Th2 cytokines, the human PBMC mRNAs differentially expressed upon the stimulation with IL-4 or IL-12, were screened by employing the differential display-polymerase chain reaction. Among a number of clones selected, DC21 was identified as a novel target gene that is regulated by IL-4 and IL-12. The DC21 gene expression was up-regulated either by IL-4 or IL-12, yet counter-regulated by co-treatment with IL-4 and IL-12. DC21 is a dendritic cell protein with an unknown function. The sequence analysis and conserved-domain search revealed that it has two AU-rich motifs in the 3'UTR, which is a target site for the regulation of mRNA stability by cytokines, and that it belongs to the N-acetyltransferase family. The induction of DC21 by IL-12 peaked around 8-12 h, and lasted until 24 h. LY294002 and SB203580 significantly suppressed the IL-12-induced DC21 gene expression, which implies that PI3K and p38/JNK are involved in the IL-12 signal transduction pathway that leads to the DC21 expression. Furthermore, tissue blot data indicated that DC21 is highly expressed in tissues with specialized-resident macrophages, such as the lung, liver, kidney, and placenta. Together, these data suggest a possible role for DC21 in the differentiation and maturation of dendritic cells regulated by IL-4 and IL-12.

• 생물정신의학
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• 제6권1호
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• pp.67-73
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• 1999

The cAMP-dependent protein kinase(PKA) is an intracellular enzyme with serine-threonine kinase activity that plays a key role in cell growth, differentiation, and apoptosis in eukaryotes. In order to understand the PKA signal transduction pathway regulating cell life cycle and identify its role, we focused on the characterization of up-/down-regulated genes by PKA using the differential display polymerase chain reaction. Seven differentially expressed sequence tags(DEST) have been obtained. Among these DESTs, 2 DESTs were homologous to the sequence of genes from BLAST search result. KC1-5 DEST that was up-regulated in A123.7 cells was highly corresponded to mouse apoptosis-related gene(MA-3) or mouse mRNA for topoisomerase inhibitor suppressed(TIS). MA-3 was induced in various types of apoptosis, specially in NGF-deprived apoptotic PC12 cells. TIS was down-regulated in the RVC lymphoma cells incubated with topoisomerase inhibitor that induces DNA strand breakages. PG1-1 DEST that was highly expressed in PC12 cells was corresponded to transposon Tn10 3'-end. Tnansposon Tn10 was up-regulated in differentiated myeloblastic ML-1 cells by 12-O-tetradecanoylphorbol-13-acetate. This study illuminates that MA-3/TIS was down-regulated by PKA activity, and transposon Tn10 was up-regulated by it.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1999년도 제13회 식물생명공학심포지움 New Approaches to Understand Gene Function in Plants and Application to Plant Biotechnology
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• pp.63-67
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• 1999

We have isolated more than a dozen cDNA clones corresponding to genes that were expressed in Arabidopsis leaves when they were kept in the dark. The nucleotide sequence analysis showed that some of the clones encoded proteins with significant homology to $\beta$-glucosidase (din2), branched-chain $\alpha$-keto acid dehydrogenase subunit E1$\beta$(din3), and another subunit E2 (din4), yeast RAD23 (din5), asparagine synthetase (din6), pre-mRNA splicing factor SRp35 (din7), phosphomannose isomerase (din9), seed imbibition protein (din10), and 2-oxoacid-dependent oxidase (din11). Accumulation of transcripts from din3,4,6 and 10 occurred rapidly after the plants were transferred to darkness. Transcripts from din2,9, and 11 could be detected only after 24 h of dark treatment. Inhibition of photo-synthesis by DCMU strongly induced the accumulation of transcripts from those genes, and application of sucrose to detached leaves suppressed the accumulation both in the dark and by DCMU. These observations indicate that expression of the genes is caused by sugar starvation resulted from the cessation of photosynthesis. We further showed that din2-encoded protein also accumulated in senescing leaves. Given these results, possible roles of din genes in leaves in the dark and senescing leaves are discussed.

• 한국어류학회지
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• 제23권1호
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• pp.1-9
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• 2011

ACP (annealing control primer)를 사용하여 DDRT (differential display reverse transcription)-PCR 방법으로 볼락의 성장단계에 따라 발현량 차이를 나타내는 DEG (differentially expressed gene)를 확보하였다. ACP 120개를 분석하여 18개월령 근육조직에서보다 6개월령 근육조직에서 발현량이 많은 DEG 16개와 6개월령 근육조직에서보다 18개월령 근육조직에서 발현량이 더 많은 DEG22개의 염기서열을 분석하였다. DEG 염기서열을 BLAST 검색한 결과, parvalbumin (PVALB) 등 18개의 유전자(PVALB, NDKB, TPM, TnI, GAPDH, CKM2, factor 2 SERF2, AMPD, TRICA, ARHGAP15, ESD, hsp70, COL1A2, GST, Midllip1, MYL1, SERCA1B, FTH1)와 69~95%의 상동성을 나타냈다. Real time PCR 분석법으로 6개월령 근육조직에서 발현량이 많은 DEG14와 PVALB 유전자의 성장단계별 발현양상을 조사한 결과, 볼락이 성장함에 따라 발현량이 감소하였으며, 특히 PVALB 유전자는 6개월령 이후에는 발현량이 극히 적었다. 6개월령 근육조직에서보다 18 개월령 근육조직에서 발현량에서 많았던 CKM2 유전자는 성장함에 따라 발현량이 계속 증가하였고, 4세 이후에는 발현량이 감소하였다. DEG의 조직특이적 발현양상을 분석한 결과, DEG14는 근육, 간, 신장, 및 비장조직에서 발현되었으며, PVALB 유전자는 근육과 신장조직에서 발현되었고, 간과 비장조직에서는 발현되지 않았다. CKM2 유전자는 근육, 신장 및 비장조직에서 발현되었고, 간 조직에서는 발현되지 않았다. PVALB 유전자의 mRNA 크기는 659 bp 이며, 110개의 아미노산으로 구성되어 있다. Parvalbumin과 CKM2 유전자는 성장속도가 빠른 어류 선발에 이용할 수 있는 분자마커 개발에 활용하고자한다.

• Radiation Oncology Journal
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• 제19권4호
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• pp.389-396
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• 2001

목적 : 임상에서 사용하는 방사선량을 조사하여 환자의 자궁경부암 세포에서 유도되는 유전자를 검색하고자 하였다. 대상 및 방법 : 자궁경부암 환자에서 방사선치료 하루 전(대조군)과 1.8 Gy 조사후 40분 지나서(조사군) 자궁경부암 조직을 생검하여 각 군에서 total RNA를 추출하였다. differential display reverse transcription-polymerase chain reaction기법(DDRT-PCR)으로 발현이 증가 또는 감소된 유전자를 탐색하였다. 발현에 변화가 있는 cDNA를 추출하고 증폭하여 얻은 클론을 reverse Northern Blot방법을 이용하여 screening하였고, Northern Blot으로 확인하였다. sequencing을 실시한 후 NCBI database를 이용하여 blast search를 하였다. 발현이 감소한 유전자를 대상으로 다른 환자에서의 발현 양상을 확인하기 위하여 방사선치료를 받는 5명의 자궁경부암 환자를 대상으로 RT-PCR로 검사 하였다. 결과 : DDRT-PCR기법을 이용하여 방사선 조사군에서 발현이 증가 혹은 감소된 18개의 cDNA band를 발견하였다. reverse northern blot을 이용한 screening에서 발현이 증가된 10개의 클론과 감소된 1개의 클론을 확인하였다. 클로닝된 cDNA 조각들은 대부분 $400\~500\;bp$ 정도 되었으며, 그중 1개의 클론은 잘 알려져 있는 chemokine receptor CXCR4 유전자와 높은 상동성을 보였으며, 4개의 클론은 Human ESTs로 확인되었고 5개의 클론은 기능이 아직 확인되지 않은 알려져 있는 염기서열로 확인되었다. 방사선에 의하여 발현이 감소한 CxCa-11 클론이 모든 환자에서 방사선치료 전 시료에서 발현을 보였으나, 방사선치료 후 시료에서는 그 발현량이 감소하거나 발현이 안되었다. 결론 : DDRT-PCR을 이용하여 임상에서 자주 사용되는 방사선량을 환자의 자궁경부암에 조사했을 때 발현되는 유전자를 확인하였다. 이러한 유전자 발현의 확인은 방사선치료과정 중에 발생하는 일련의 기전들을 이해하는데 도움이 되리라 생각된다.

• Asian-Australasian Journal of Animal Sciences
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• 제20권9호
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• pp.1334-1341
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• 2007

This study identified hepatic differentially expressed genes (DEGs) affecting the marbling of muscle. Most dietary nutrients bypass the liver and produce plasma lipoproteins. These plasma lipoproteins transport free fatty acids to the target tissue, adipose tissue and muscle. We examined hepatic genes differentially expressed in a differential-display reverse transcription-polymerase chain reaction (ddRT-PCR) analysis comparing high- and low-marbled Hanwoo steers. Using 60 arbitrary primers, we found 13 candidate genes that were upregulated and five candidate genes that were downregulated in the livers of high-marbled Hanwoo steers compared to low-marbled individuals. A BLAST search for the 18 DEGs revealed that 14 were well characterized, while four were not annotated. We examined four DEGs: ATP synthase F0, complement component CD, insulin-like growth factor binding protein-3 (IGFBP3) and phosphatidylethanolamine binding protein (PEBP). Of these, only two genes (complement component CD and IGFBP3) were differentially expressed at p<0.05 between the livers of high- and low-marbled individuals. The mean mRNA levels of the PEBP and ATP synthase F0 genes did not differ significantly between the livers of high- and low-marbled individuals. Moreover, these DEGs showed very high inter-individual variation in expression. These informative DEGs were assigned to the bovine chromosome in a BLAST search of MS marker subsets and the bovine genome sequence. Genes related to energy metabolism (ATP synthase F0, ketohexokinase, electron-transfer flavoprotein-ubiquinone oxidoreductase and NADH hydrogenase) were assigned to BTA 1, 11, 17, and 22, respectively. Syntaxin, IGFBP3, decorin, the bax inhibitor gene and the PEBP gene were assigned to BTA 3, 4, 5, 5, and 17, respectively. In this study, the in silico physical maps provided information on the specific location of candidate genes associated with economic traits in cattle.

• 한국수정란이식학회지
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• 제33권4호
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• pp.195-203
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• 2018

Interferon-tau (IFNT) is known as a major conceptus protein that signals the process of maternal recognition of pregnancy in ruminants. Also, multiple interferon genes exist in cattle, However, molecular mechanisms of these bovine IFNT (bIFNT) genes whose expressions are limited have not been characterized. We and others have observed that expression levels of bovine subtype IFNT genes in the tissues of ruminants; thus, bIFNT1 and other new type I (bIFNTc1/c2/c3) gene co-exist during the early stages of conceptus development and non-trophoblast cells. Its genes transcription could be regulated through CDX2 and ETS2 and JUN and/or cAMP-response element binding protein (CREB)-binding protein (CREBBP) expression, a transcription factor implicated in the control of cell differentiation in the trophectoderm. Bovine ear-derived fibroblast cells, were co-transfected with luciferase reporter constructs carrying upstream (positions -1000 to +51) regions of bIFNT1 and other new type I gene and various transcription factor expression plasmids. Compared to each - 1kb-bIFNT1/c1/c2/c3-Luc increased when this constructs were co-transfected with CDX2, ETS2, JUN and/or CREBBP. Also, Its genes was had very effect on activity by CDX2, either alone or with the other transcription factors, markedly increased luciferase activity. However, the degree of transcriptional activation of the bIFNTc1 gene was not similar to that bIFNT1/c2/c3 gene by expression plasmid. Furthermore, Sequence analyses also revealed that the expression levels of bIFNT1/c2/c3 gene mRNAs expression were highest on day 17, 20 and 22 trophoblast and, Madin-Darby bovine kidney (MDBK), Bovine ear-derived fibroblast (EF), and endometrium (Endo) non-trophoblast cells. But, bIFNTc1 mRNA had not same expression level, bIFNTc1 lowest levels than those of IFNT1/c2/c3 gene in both trophoblast and non-trophoblast cells. These results demonstrate that bovine subtype bIFNT genes display differential, in the trophoblast and non-trophoblast cells.

• Asian-Australasian Journal of Animal Sciences
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• 제27권1호
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• pp.10-18
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• 2014

The objective of this study was to investigate the correlation between cattle breeds and deposit of adipose tissues in different positions and the gene expressions of peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$), fatty acid synthase (FASN), and Acyl-CoA dehydrogenase (ACADM), which are associated with lipid metabolism and are valuable for understanding the physiology in fat depot and meat quality. Yanbian yellow cattle and Yan yellow cattle reared under the same conditions display different fat proportions in the carcass. To understand this difference, the expression of $PPAR{\gamma}$, FASN, and ACADM in different adipose tissues and longissimus dorsi muscle (LD) in these two breeds were analyzed using the Real-time quantitative polymerase chain reaction method (qRT-PCR). The result showed that $PPAR{\gamma}$ gene expression was significantly higher in adipose tissue than in LD in both breeds. $PPAR{\gamma}$ expression was also higher in abdominal fat, in perirenal fat than in the subcutaneous fat (p<0.05) in Yanbian yellow cattle, and was significantly higher in subcutaneous fat in Yan yellow cattle than that in Yanbian yellow cattle. On the other hand, FASN mRNA expression levels in subcutaneous fat and abdominal fat in Yan yellow cattle were significantly higher than that in Yanbian yellow cattle. Interestingly, ACADM gene shows greater fold changes in LD than in adipose tissues in Yan yellow cattle. Furthermore, the expressions of these three genes in lung, colon, kidney, liver and heart of Yanbian yellow cattle and Yan yellow cattle were also investigated. The results showed that the highest expression levels of $PPAR{\gamma}$ and FASN genes were detected in the lung in both breeds. The expression of ACADM gene in kidney and liver were higher than that in other organs in Yanbian yellow cattle, the comparison was not statistically significant in Yan yellow cattle.

• The Korean Journal of Physiology and Pharmacology
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• 제9권5호
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• pp.275-282
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• 2005

By using differential display, we identified one of the genes encoding the multi-subunit complex protein V-ATPase, c subunit gene (ATP6L), and showed alterations of the gene expression by oxidative stresses. Expression of the ATP6L gene in Neuro-2A cells was increased by the treatment with $H_2O_2$ and incubation in hypoxic chamber, implying that the expression of the ATP6L gene is regulated by oxidative stresses. To examine mechanisms involved in the regulation of the gene expression by oxidative stresses, the transcriptional activity of the rat ATP6L promoter was studied. Transcription initiation site was determined by primer extension analysis and DNA sequencing, and promoter of the rat ATP6L and its deletion clones were constructed in reporter assay vector. Significant changes of the promoter activities in Neuro-2A cells were observed in two regions within the proximal 1 kbp promoter, and one containing a suppressor was in -195 to -220, which contains GC box that is activated by binding of Sp1 protein. The suppression of promoter activity was lost in mutants of the GC box. We confirmed by electrophoretic mobility shift and supershift assays that Sp1 protein specifically binds to the GC box. The promoter activity was not changed by the $H_2O_2$ treatment and incubation in hypoxic chamber, however, $H_2O_2$ increased the stability of ATP6L mRNA. These data suggest that the expression of the ATP6L gene by oxidative stresses is regulated at posttranscriptional level, whereas the GC box is important in basal activities of the promoter.

• 한국잠사곤충학회지
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• 제51권2호
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• pp.211-217
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• 2013

일반적으로 누에나방의 평균 생존시수는 암컷과 수컷이 각각 8일과 5일로 알려져 있다. 그러나 J037 품종의 성충 수명은 상당히 길며, 반대로 Daizo(sdi) 품종은 상당히 짧은 성충 수명 특성을 갖고 있다. 이러한 누에 품종 간 성충 수명의 차이는 유전적 차이에 따른 현상으로 추정되어 왔다. 본 연구에서는 장단명누에 품종의 발현유전자를 분석함으로써 누에 수명 관련 유전자를 탐색하여 그 특성을 분석하고자 하였다. 우선, 장명 수나방 mRNA를 사용하여 cDNA 유전자은행을 제작하고, 제작된 유전자은행으로부터 2,688개 클론을 무작위 선발한 후 장단명 수나방 cDNA를 각각의 탐침으로 차별화선별을 수행하였다. 본 연구에서 차별화 발현하는 193개의 클론을 선발하여 EST 상동성 분석을 통해 최종적으로 154개의 J037 수나방 수명 관련 독립유전자를 선발할 수 있었는데, 장명 탐침에서 과발현하는 124개와 단명 탐침에서 과발현하는 30개로 구성된다. 154개 독립유전자 중 가장 많은 발현빈도수를 보인 유전자는 cytochrome oxidase subunit-1으로서 모두 9회로 확인되었고, 현재까지 기능에 관해 알려진 바 없는 1-50번 독립유전자가 5회로 두 번째로 높은 발현빈도를 보였다. 154개 선발 독립유전자를 기능별로 분류한 결과, unclassified protein군에 가장 많은 24%의 독립유전자가 포함되어 있었다. 본 연구에서 두 번째로 발현빈도가 높은 클론(ID;1-50)의 염기서열 및 아미노산 서열을 분석하였는데, 1-50번 유전자는 전체 1,523 bp 염기로 구성되어 있으며, 723개 염기쌍이 240개의 아미노산을 coding 하고 있었다. 본 연구에서 선발된 주요 수명 관련 유전자는 국제유전자은행 EST database에 등록하였다.