• 제목/요약/키워드: gene discovery

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정상, 낭종 및 법랑아세포종 세포에서의 유전자 발현 차이 분석 (ANALYSIS OF DIFFERENTIAL GENE EXPRESSION IN NORMAL, CYST AND AMELOBLASTOMA CELLS)

  • 양철희;백병주;양연미;김재곤
    • 대한소아치과학회지
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    • 제32권1호
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    • pp.75-88
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    • 2005
  • 법랑아세포종은 1868년에 처음 보고된 이래 명칭, 발생기전, 분류 그리고 치료 방법 등에 관하여 수 많은 논란이 있어 왔는데 이는 법랑세포종이 양성종양임에도 불구하고 종양자체의 진행이 파괴적이고, 외과적 처치를 한 후에도 재발이 잘되며, 흔하지는 않지만 악성종양과 유사하게 전이를 보이는 등 독특한 특성을 지니고 있기 때문이다. 정상세포와 암 세포 간에 차이를 보이는 유전자 혹은 정상세포에서 변형이 일어날 때 특이적으로 발현하는 유전의 분리 및 분석하는 것은 암세포의 생성과정을 이해하는데 있어서 중요한 열쇠를 제공할 수 있다. 이에 본 연구는 RNA differential display 방법 중 재연성과 반복성이 개선된 Ordered differential display(ODD)RT-PCR과 보다 개선된 $GeneFishing^{TM}$기술을 이용하여 악성과 양성종양 사이의 유전자 발현의 차이를 조사하고, 특이 유전자의 profile을 확보하고자 하였다. $GeneFishing^{TM}$기술과 RT-PCR을 수행한 결과 nasopharyngeal carcinoma gene을 제외한 9개의 유전자는 악성에서 특이적으로 발현되는 것을 확인하였다. 따라서 $GeneFishing^{TM}$을 이용하면 각 시료간의 mRNA 상에서 발현차이를 보이는 DEG를 비교 분석하면 암관련 유전자, 항생제 태성 유전자, 그리고 분화 관련 유전자들에 대한 연구가 용이하게 수행할 수 있을 것으로 생각된다.

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EST-based Identification of Genes Expressed in the Muscle of Olive Flounder, Paralichthys olivaceus

  • Park, Eun-Mi;Kim, Young-Ok;Nam, Bo-Hye;Kong, Hee Jeong;Kim, Woo-Jin;Lee, Sang-Jun;Choi, Tae-Jin
    • 한국해양바이오학회지
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    • 제2권3호
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    • pp.168-173
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    • 2007
  • of expressed sequence tags (ESTs) is an efficient approach for gene discovery, expression profiling, and development of resources useful for functional genomics. To analyze the transcriptome of olive flounder, Paralichthys olivaceus, we have conducted EST analysis using cDNA libraries made from muscle of P. olivaceus. Redundant ESTs were assembled into overlapping contigs by using the assembly program ICAtools software. We found that the 221 ESTs were composed of 21 clusters and 35 singletons, suggesting that the overall redundancy of the library was 74.7%. Of the 221 clones, 218 clones (98.6%) were identified as known genes by BLAST searches and 3 clones (1.4%) did not match to any previously described genes. Based on major functions of their encoded proteins, the identified clones were classified into 13 broad categories. Sequence analysis of the ESTs revealed the presence of microsatellite-containing genes which may be valuable for further gene mapping studies. This study contributes to the identification of many EST clones that could be useful for genetics and developmental biology of olive flounder.

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Emerging Genomics Technologies in Nutritional Sciences: Applications to obesity and hypertension research

  • Mouss, Naima-Moustaid;Sumithra Urs;Kim, Suyeon;Heo, Young-Ran
    • 한국영양학회:학술대회논문집
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    • 한국영양학회 2002년도 춘계학술대회
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    • pp.29-41
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    • 2002
  • While the sequencing of several genomes was underway, several advanced techniques in genetics, molecular biology and protein chemistry emerged. Within the nutritional sciences, while the focus on nutrition education, epidemiology and public health aspects remains essential; it is crucial to incorporate the new advances in gene and protein discovery in nutritional studies. Nutrition is a discipline that has always integrated social, biochemical and physiological sciences from the studies at the molecule level to studies at the population level. For this reason, nutritionists are in a prime position to readily incorporate the current genomics approaches in nutrition research, All the available analytical techniques can and should be used in modern nutritional sciences. These include genetics, genomics, proteomics and metabolomics which also require integration and use of bioinformatics and computational methods for data analysis and management. These applications will be briefly reviewed with a primary focus on what the genomics and genetics approaches offer to nutritionists. We will use one of our research focus areas to illustrate uses of some of these applications in obesity-hypertension research. Our central hypothesis is that adipose tissue is an endocrine organ that plays a major role in obesity and related hypertension. We are primarily studying the renin angiotensin system (RAS). We provide evidence from our own studies and others for the paracrine as well as endocrine role of adipocyte-derived angiotensin II in adipocyte gene expression, adiposity and blood pressure regulation. Both cell culture studies as well as knockout and transgenic mice models are used to test our hypothesis. Genomics and proteomics technologies are currently developed to complement our physiological and molecular studies on the RAS and for a fine analysis of this system and its function in health and disease.

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Novel Bacterial Surface Display System Based on the Escherichia coli Protein MipA

  • Han, Mee-Jung
    • Journal of Microbiology and Biotechnology
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    • 제30권7호
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    • pp.1097-1103
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    • 2020
  • Bacterial surface display systems have been developed for various applications in biotechnology and industry. Particularly, the discovery and design of anchoring motifs is highly important for the successful display of a target protein or peptide on the surface of bacteria. In this study, an efficient display system on Escherichia coli was developed using novel anchoring motifs designed from the E. coli mipA gene. Using the C-terminal fusion system of an industrial enzyme, Pseudomonas fluorescens lipase, six possible fusion sites, V140, V176, K179, V226, V232, and K234, which were truncated from the C-terminal end of the mipA gene (MV140, MV176, MV179, MV226, MV232, and MV234) were examined. The whole-cell lipase activities showed that MV140 was the best among the six anchoring motifs. Furthermore, the lipase activity obtained using MV140 as the anchoring motif was approximately 20-fold higher than that of the previous anchoring motifs FadL and OprF but slightly higher than that of YiaTR232. Western blotting and confocal microscopy further confirmed the localization of the fusion lipase displayed on the E. coli surface using the truncated MV140. Additionally the MV140 motif could be used for successfully displaying another industrial enzyme, α-amylase from Bacillus subtilis. These results showed that the fusion proteins using the MV140 motif had notably high enzyme activities and did not exert any adverse effects on either cell growth or outer membrane integrity. Thus, this study shows that MipA can be used as a novel anchoring motif for more efficient bacterial surface display in the biotechnological and industrial fields.

Circulating microRNA expression profiling in young obese Korean women

  • Choi, Won Hee;Ahn, Jiyun;Um, Min Young;Jung, Chang Hwa;Jung, Sung Eun;Ha, Tae Youl
    • Nutrition Research and Practice
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    • 제14권4호
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    • pp.412-422
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    • 2020
  • BACKGROUND/OBJECTIVES: This study investigates correlations between circulating microRNAs (miRNAs) and obesity-related parameters among young women (aged 20-30 years old) in Korea. SUBJECTS/METHODS: We analyzed TaqMan low density arrays (TLDAs) of circulating miRNAs in 9 lean (body mass index [BMI] < 25 kg/㎡) and 15 obese (BMI > 25 kg/㎡) women. We also performed gene ontology (GO) analyses of the biological functions of predicted miRNA target genes, and clustered the results using the database for annotation, visualization and integrated discovery. RESULTS: The TLDA cards contain 754 human miRNAs; of these, the levels of 8 circulating miRNAs significantly declined (> 2-fold) in obese subjects compared with those in lean subjects, including miR-1227, miR-144-5p, miR-192, miR-320, miR-320b, miR-484, miR-324-3p, and miR-378. Among them, miR-484 and miR-378 displayed the most significant inverse correlations with BMI (miR-484, r = -0.5484, P = 0.0056; miR-378, r = -0.5538, P = 0.0050) and visceral fat content (miR-484, r = -0.6141, P = 0.0014; miR-378, r = -0.6090, P = 0.0017). GO analysis indicated that genes targeted by miR-484 and miR-378 had major roles in carbohydrate and lipid metabolism. CONCLUSION: Our result showed the differentially expressed circulating miRNAs in obese subjects compared to lean subjects. Although the mechanistic study to reveal the causal role of miRNAs remains, these miRNAs may be novel biomarkers for obesity.

Emerging Genomics Technologies in Nutritional Sciences : Applications to Obesity and Hypertension Research

  • Moustaid-Moussa;Sumithra Urs;Kim, Suyeon;Heo, Young-Ran
    • 한국영양학회:학술대회논문집
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    • 한국영양학회 2002년도 춘계 심포지움초록
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    • pp.598-603
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    • 2002
  • While the sequencing of several genomes was underway, several advanced techniques in genetics, molecular biology and protein chemistry emerged. Within the notritional sciences, while the focus on nutrition education, epidemiology and public health aspects remains essential; it is crucial to incorporate the new advances in gene and protein discovery in nutritional studies. Nutrition is a discipline that has always integrated social, biochemical and physiological sciences from the studies at the molecule level to studies at the population level. for this reason, nutritionists are in a prime position to readily incorporate the current genomics approaches in nutrition research. All the available analytical techniques can and should be used in modem nutritional sciences. These include genetics, genomics, proteomics and metabolomics which also require integration and use of bioinformatics and computational methods for data analysis and management. These applications will be briefly reviewed with a primary focus on what the genomics and genetics approaches offer to nutritionists. We will use one of our research focus areas to illustrate uses of some of these applications in obesity-hypertension research. Our central hypothesis is that adipose tissue is an endocrine organ that plays a major role in obesity and related hypertension. We are primarily studying the renin angiotensin system (RAS). We provide evidence from our own studies and others for the paracrine as well as endocrine role of adipocyte-derived angiotensin II in adipocyte gene expression, adiposity and blood pressure regulation. Both cell culture studies as well as knockout and transgenic mice models are used to test our hypothesis. Genomics and proteomics technologies are currently developed to complement our physiological and molecular studies on the RAS and for a fine analysis of this system and its function in health and disease.

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Plant Phenolics Ferulic Acid and P-Coumaric Acid Inhibit Colorectal Cancer Cell Proliferation through EGFR Down-Regulation

  • Roy, Nabarun;Narayanankutty, Arunaksharan;Nazeem, PA;Valsalan, Ravisankar;Babu, TD;Mathew, Deepu
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권8호
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    • pp.4019-4023
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    • 2016
  • Background: Colorectal cancer (CRC) or bowel cancer is one of the most important cancer diseases, needing serious attention. The cell surface receptor gene human epidermal growth factor receptor (EGFR) may have an important role in provoking CRC. In this pharmaceutical era, it is always attempted to identify plant-based drugs for cancer, which will have less side effects for human body, unlike the chemically synthesized marketed drugs having serious side effects. So, in this study the authors tried to assess the activity of two important plant compounds, ferulic acid (FA) and p-coumaric acid (pCA), on CRC. Materials and Methods: FA and pCA were tested for their cytotoxic effects on the human CRC cell line HCT 15 and also checked for the level of gene expression of EGFR by real time PCR analysis. Positive results were confirmed by in silico molecular docking studies using Discovery Studio (DS) 4.0. The drug parallel features of the same compounds were also assessed in silico. Results: Cytotoxicity experiments revealed that both the compounds were efficient in killing CRC cells on a controlled concentration basis. In addition, EGFR expression was down-regulated in the presence of the compounds. Docking studies unveiled that both the compounds were able to inhibit EGFR at its active site. Pharmacokinetic analysis of these compounds opened up their drug like behaviour. Conclusions: The findings of this study emphasize the importance of plant compounds for targeting diseases like CRC.

Differential expression of microRNAs in the saliva of patients with aggressive periodontitis: a pilot study of potential biomarkers for aggressive periodontitis

  • Lee, Nam-Hun;Lee, Eunhye;Kim, Young-Sung;Kim, Won-Kyung;Lee, Young-Kyoo;Kim, Su-Hwan
    • Journal of Periodontal and Implant Science
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    • 제50권5호
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    • pp.281-290
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    • 2020
  • Purpose: The aim of this study was to compare microRNA (miRNA) gene expression in saliva using miRNA polymerase chain reaction (PCR) arrays in healthy and aggressive periodontitis (AP) patients. Methods: PCR arrays of 84 miRNAs related to the human inflammatory response and autoimmunity from the saliva samples of 4 patients with AP and 4 healthy controls were performed. The functions and diseases related to the miRNAs were obtained using TAM 2.0. Experimentally validated targets of differentially expressed miRNAs were obtained from mirTarBase. Gene ontology terms and pathways were analyzed using ConsensusPathDB. Results: Four downregulated miRNAs (hsa-let-7a-5p, hsa-let-7f-5p, hsa-miR-181b-5p, and hsa-miR-23b-3p) were identified in patients with AP. These miRNAs are associated with cell death and innate immunity, and they target genes associated with osteoclast development and function. Conclusions: This study is the first analysis of miRNAs in the saliva of patients with AP. Identifying discriminatory human salivary miRNA biomarkers reflective of periodontal disease in a non-invasive screening assay is crucial for the development of salivary diagnostics. These data provide a first step towards the discovery of key salivary miRNA biomarkers for AP.

Transcriptional Alteration of p53 Related Processes As a Key Factor for Skeletal Muscle Characteristics in Sus scrofa

  • Kim, Seung-Soo;Kim, Jung-Rok;Moon, Jin-Kyoo;Choi, Bong-Hwan;Kim, Tae-Hun;Kim, Kwan-Suk;Kim, Jong-Joo;Lee, Cheol-Koo
    • Molecules and Cells
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    • 제28권6호
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    • pp.565-573
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    • 2009
  • The pig could be a useful model to characterize molecular aspects determining several delicate phenotypes because they have been bred for those characteristics. The Korean native pig (KNP) is a regional breed in Korea that was characterized by relatively high intramuscular fat content and reddish meat color compared to other western breeds such as Yorkshire (YS). YS grew faster and contained more lean muscle than KNP. We compared the KNP to Yorksire to find molecular clues determining muscle characteristics. The comparison of skeletal gene expression profiles between these two breeds showed molecular differences in muscle. We found 82 differentially expressed genes (DEGs) defined by fold change (more than 1.5 fold difference) and statistical significance (within 5% of false discovery rate). Functional analyses of these DEGs indicated up-regulation of most genes involved in cell cycle arrest, down-regulation of most genes involved in cellular differentiation and its inhibition, down-regulation of most genes encoding component of muscular-structural system, and up-regulation of most genes involved in diverse metabolism in KNP. Especially, DEGs in above-mentioned categories included a large number of genes encoding proteins directly or indirectly involved in p53 pathway. Our results indicated a possible role of p53 to determine muscle characteristics between these two breeds.

Reversal of Multidrug Resistance in Mouse Lymphoma Cells by Extracts and Flavonoids from Pistacia integerrima

  • Rauf, Abdur;Uddin, Ghias;Raza, Muslim;Ahmad, Bashir;Jehan, Noor;Siddiqui, Bina S;Molnar, Joseph;Csonka, Akos;Szabo, Diana
    • Asian Pacific Journal of Cancer Prevention
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    • 제17권1호
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    • pp.51-55
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    • 2016
  • Phytochemical investigation of Pistacia integerrima has highlighted isolation of two known compounds naringenin (1) and dihydrokaempferol (2). A crude extract and these isolated compounds were here evaluated for their effects on reversion of multidrug resistance (MDR) mediated by P-glycoprotein (P-gp). The multidrug resistance P-glycoprotein is a target for chemotherapeutic drugs from cancer cells. In the present study rhodamine-123 exclusion screening test on human mdr1 gene transfected mouse gene transfected L5178 and L5178Y mouse T-cell lymphoma cells showed excellent MDR reversing effects in a dose dependent manner. In-silico molecular docking investigations demonstrated a common binding site for Rhodamine123, and compounds naringenin and dihydrokaempferol. Our results showed that the relative docking energies estimated by docking softwares were in satisfactory correlation with the experimental activities. Preliminary interaction profile of P-gp docked complexes were also analysed in order to understand the nature of binding modes of these compounds. Our computational investigation suggested that the compounds interactions with the hydrophobic pocket of P-gp are mainly related to the inhibitory activity. Moreover this study s a platform for the discovery of novel natural compounds from herbal origin, as inhibitor molecules against the P-glycoprotein for the treatment of cancer.