• 제목/요약/키워드: protein arrays

검색결과 38건 처리시간 0.046초

Lack of Increased P15INK4B Protein Expression in Basal Cell Carcinomas

  • Moad, Ahmed Ismail Hassan;Tan, Mei Lan;Kaur, Gurjeet;Mabruk, Mohamed
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권12호
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    • pp.6239-6244
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    • 2012
  • Background: The basal cell carcinoma (BCC) is the most common non-melanoma skin cancer (NMSK). BCC might develop because of the faulty cell cycle arrest. $p15^{INK4b}$ is a tumor suppressor gene, involved in cell cycle arrest and inactivated in most human cancers. The role of $p15^{INK4b}$ protein expression in the genesis of BCC is as yet unknown. In a previous study we showed the absence of $p15^{INK4b}$ expression in the majority of tissue microarray cores of cutaneous squamous cell carcinoma (SCCs), another type of non-melanoma skin cancer, indicating that $p15^{INK4b}$ could possibly be involved in the pathogenesis of cutaneous SCC. The aim of this study was to investigate $p15^{INK4b}$ protein expression in BCCs. Materials and Method: Protein expression of $p15^{INK4b}$ in 35 cases of BCC tissue arrays and 19 cases of normal human skin tissue was studied using an immunohistochemical approach. Results: The expression of $p15^{INK4b}$ was not significantly different in the BCC cases as compared with normal human skin (p=0.356; p>0.05). In addition, there were no significant relationship between clinicopathologic variables of patients (age and sex) and $p15^{INK4b}$ protein expression. Conclusions: Our finding may indicate that $p15^{INK4b}$ protein expression does not play a role in the genesis of BCC.

Expression Profiles of Apoptosis Genes in Mammary Epithelial Cells

  • Seol, Myung Bok;Bong, Jin Jong;Baik, Myunggi
    • Molecules and Cells
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    • 제20권1호
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    • pp.97-104
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    • 2005
  • To investigate apoptosis in HC11 mammary epithelial cells, we compared the gene expression profiles of actively growing and serum-starved apoptotic cells using a mouse apoptosis gene array and $^{33}P$-labeled cDNA prepared from the RNA of the two cultures. Analysis of the arrays showed that expression of several genes such as clusterin, secreted frizzled related protein mRNA (sFRP-1), CREB-binding protein (CBP), and others was higher in the apoptotic cells whereas expression of certain genes including survivin, cell division cycle 2 homolog A (CDC2), and cyclin A was lower. These expression patterns were confirmed by RT-PCR and/or Northern analyses. We compared the expression of some of these genes in the mouse mammary gland under various physiological conditions. The expression levels of genes (clusterin, CBP, and M6P-R) up-regulated in apoptotic conditions were higher at involution than during lactation. On the other hand, genes (Pin, CDC2) downregulated in apoptotic conditions were relatively highly expressed in virgin and pregnant mice. We conclude that certain genes such as clusterin, sFRP-1, GAS1 and CBP are induced in apoptotic mammary epithelial cells, and others are repressed. Moreover, the apoptosis array is an efficient technique for comparing gene expression profiles in different states of the same cell type.

크로스 링크된 단백질 서브시퀀스를 찾는 알고리즘 (Algorithm for identifying cross-linked protein subsequences)

  • 김성권
    • 한국정보과학회논문지:시스템및이론
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    • 제29권9호
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    • pp.514-519
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    • 2002
  • 단백질의 구조를 예측하는 과정에 사용될 수 있는 다음 문제를 고려한다. 길이가 n이고 원소가 모두 양수인 두 배열 A, B와 양수 M이 주어질 때, A[i]+…A[j]+B[k]+…B[ι]=M이 되는 부배열 쌍 A[i]+…A[j],$1{\leq}i{\leq}j{\leq}n$과 B[k], …, B[l], $1{\leq}k{\leq}l{\leq}n$을 모두 찾으시오. 본 논문에서는 이 문제를 $Ο(n^2log n+K)$ 시간에 Ο(n) 메모리를 사용하여 해결하는 알고리즘을 제시한다. 단, K는 찾은 부배열 쌍의 수이다. 기존의 결과는$Ο(n^2log +Klog n)$ 시간과 Ο(n) 메모리였다.

Identification of Sugar-Responsive Genes and Discovery of the New Functions in Plant Cell Wall

  • 이은정
    • 한국작물학회:학술대회논문집
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    • 한국작물학회 2007년도 춘계학술발표회
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    • pp.65-73
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    • 2007
  • The objective of this study is to understand how regulatory mechanisms respond to sugar status for more efficient carbon utilization and source-sink regulation in plants. So, we need to identify and characterize many components of sugar-response pathways for a better understanding of sugar responses. For this end, genes responding change of sugar status were screened using Arabidpsis cDNA arrays, and confirmed thirty-six genes to be regulated by sucrose supply in detached leaves by RNA blot analysis. Eleven of them encoding proteins for amino acid metabolism and carbohydrate metabolism were repressed by sugars. The remaining genes induced by sugar supply were for protein synthesis including ribosomal proteins and elongation factors. Among them, I focused on three hydrolase genes encoding putative $\beta$-galactosidase, $\beta$-xylosidase, and $\beta$-glucosidase that were transcriptionally induced in sugar starvation. Homology search indicated that these enzymes were involved in hydrolysis of cell wall polysaccharides. In addition to my results, recent transcriptome analysis suggested multiple genes for cell wall degradation were induced by sugar starvation. Thus, I hypothesized that enzyme for cell wall degradation were synthesized and secreted to hydrolyze cell wall polysaccharides producing carbon source under sugar-starved conditions. In fact, the enzymatic activities of these three enzymes increased in culture medium of Arabidopsis suspension cells under sugar starvation. The $\beta$-galactosidase encoded by At5g56870 was identified as a secretory protein in culture medium of suspension cells by mass spectrometry analysis. This protein was specifically detected under sugar-starved condition with a specific antibody. Induction of these genes was repressed in suspension cells grown with galactose, xylose and glucose as well as with sucrose. In planta, expression of the genes and protein accumulation were detected when photosynthesis was inhibited. Glycosyl hydrolase activity against galactan also increased during sugar starvation. Further, contents of cell wall polysaccharides especially pectin and hemicellulose were markedly decreased associating with sugar starvation in detached leaves. The amount of monosaccharide in pectin and hemicellulose in detached leaves decreased in response to sugar starvation. These results supported my idea that cell wall has one of function to supply carbon source in addition to determination of cell shape and physical support of plant bodies.

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Kinesin superfamily KIF1A와 결합하는 미세소관 불안정화 단백질 SCG10의 규명 (SCG10, a Microtubule-Destabilizing Factor, Interacts Directly with Kinesin Superfamily KIF1A Protein in Brain)

  • 문일수;석대현
    • 생명과학회지
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    • 제19권7호
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    • pp.859-865
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    • 2009
  • 미세소관은 세포골격단백질의 중요한 구성 단백질로 축삭돌기 내에서는 세포막 방향으로 정렬되어 있다. Kinesin superfamily (KIFs)는 세포 내에서 미세소관을 따라 세포 내 소포들을 운반하는 분자 자동차 (molecular motor) 단백질이다. 본 연구에서 우리는 효모 two-hybrid system을 사용하여 KIF1A의 coiled-coil 영역과 결합하는 단백질로 미세소관 불안정화 요소인 SCG10 단백질을 분리하였다. SCG10은 KIFs에서 KIF1A와만 특이적으로 결한 하며, KIF1A의 400에서 820아미노산 부위가 SCG10과의 결합에 필수적임을 효모 two-hybrid assay로 확인하였다. 또한 SCG10의 coiled-coil영역은 KIF1A와의 결합에 필수영역임을 확인하였으며 단백질간의 결합은 Glutathione S-transferase pull-down assay를 통하여 확인하였다. 생쥐의 뇌 파쇄액에 SCG10항체로 면역침강을 행하여 KIF1A를 확인한 결과KIF1A는 SCG10과 특이적으로 같이 침강하였다. 이러한 결과들은 KIF1A는 SCG10와 결합하여 SCG10이 포함된 소포를 미세소관을 따라 이동시킴을 시사한다.

Targeting Renal Cell Carcinoma with Gambogic Acid in Combination with Sunitinib in Vitro and in Vivo

  • Jiang, Xiao-Liang;Zhang, Yao;Luo, Chun-Li;Wu, Xiao-Hou
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권12호
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    • pp.6463-6468
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    • 2012
  • Purpose: To evaluated the effect of the gambogic acid (GA), one of the effective components of Garcinia, in combination with a new multi-targeted oral medication, sunitinib (SU) on renal cancer cell proliferation in vitro and on tumor growth in vivo. Methods: After treatment with GA or SU, either alone or in combination, MTT and FACS analysis were used to examine cell viability and cycle distribution of the renal carcinoma cell lines 786-0 and Caki-1. Western blotting was employed to examine the expression of proteins related to the cell cycle and vascular formation. Furthermore, a xenograft model was applied to study the antitumor efficacy of SU or GA alone or in combination, with immunohistochemistry to detect expression of proteins related to xenograft growth and angiogenesis. Western blotting was used to examine NF-${\kappa}B$ signaling pathway elements in xenografts. Results: Treatment of 786-0 and Caki-1 cells with GA or SU resulted in decreased tumor cell proliferation, especially with joint use. Cells accumulated more strongly in the sub-G1 phase after joint treatment with GA and SU than treatment of GA and SU alone. Western blotting arrays showed 1 protein significantly upregulated, 2 proteins downregulated, and 2 proteins unchanged. Moreover, combined use of GA and SU inhibited the growth and angiogenesis of xenografts generated from Caki-1 significantly. Immunohistochemistry arrays showed downregulation of the expression of proteins promoting xenograft growth and angiogenesis, and Western blotting showed inhibition of the NF-${\kappa}B$ signaling pathway after treatment by GA alone and in combination with SU in xenografts. Conclusions: Our results show that the joint use of GA and SU can provide greater antitumor efficacy compared to either drug alone and thus may offer a new treatment strategy for renal cell carcinoma.

남극 지의류에서 분리한 Caballeronia sordidicola균주 PAMC 26510의 유전체 서열 분석 (Genome sequence of Caballeronia sordidicola strain PAMC 26510 isolated from Psoroma sp., an Antarctic lichen)

  • 양정안;홍순규;오현명
    • 미생물학회지
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    • 제53권2호
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    • pp.137-140
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    • 2017
  • Caballeronia sordidicola 균주 PAMC 26510은 남극 킹조지섬의 바톤반도에서 채집된 지의류인 Psoroma sp.에서 분리되었다. 균주 PAMC 26510의 초안 유전체 서열은 224개의 콘티그로 이루어졌으며, 총 7,872,143 염기쌍은 59.7% G+C 함량을 나타냈다. 유전체는 7,580개의 단백질 유전자, 6개의 rRNA 유전자와 46개의 tRNA 유전자를 포함하였다. 균주 PAMC 26510는 이전에 연구한 Caballeronia sordidicola에 속하는 북극 균주와 마찬가지로 대사능력에 있어서 광범위한 능력을 가지고 있는 것으로 여겨진다. PAMC 26510의 초안 유전체는 6개의 CRISPR arrays를 각각의 콘티그 상에 가지고 있으며, 이중 두 개의 콘티그 상에 CRISPR-관련 유전자가 연결되어 있었다.

Discovering Novel Genes of poultry in Genomic Era

  • S.K. Kang;Lee, B.C.;J.M. Lim;J.Y. Han;W.S. Hwang
    • 한국가금학회지
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    • 제28권2호
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    • pp.143-153
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    • 2001
  • Using bioinformatic tools for searching the massive genome databases, it is possible to Identify new genes in few minutes for initial discoveries based on evolutionary conservation, domain homology, and tissue expression patterns, followed by further verification and characterization using the bench-top works. The development of high-density two-dimensional arrays has allowed the analysis of the expression of thousands of genes simultaneously in the humans, mice, rats, yeast, and bacteria to elucidate the genes and pathways involved in physiological processes. In addition, rapid and automated protein identification is being achieved by searching protein and nucleotide sequence databases directly with data generated from mass spectrometry. Recently, analysis at the bio-chemical level such as biochemical screening and metabolic profiling (Biochemical genomics) has been introduced as an additional approach for categorical assignment of gene function. To make advantage of recent achievements in computational approaches for facilitated gene discoveries in the avian model, chicken expression sequence tags (ESTs) have been reported and deposited in the international databases. By searching EST databases, a chicken heparanase gene was identified and functionally confirmed by subsequent experiments. Using combination of sub-tractive hybridization assay and Genbank database searches, a chicken heme -binding protein family (cSOUL/HBP) was isolated in the retina and pineal gland of domestic chicken and verified by Northern blot analysis. Microarrays have identified several host genes whose expression levels are elevated following infection of chicken embryo fibroblasts (CEF) with Marek's disease virus (MDV). The ongoing process of chicken genome projects and new discoveries and breakthroughs in genomics and proteomics will no doubt reveal new and exciting information and advances in the avian research.

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Variation of Anthocyanin and Protein Contents in Glycine max L. (Merr) (Soybean) Germplasms from Korea

  • Choi, Yu Mi;Lee, Sukyeung;Hyun, Do-Yoon;Ko, Ho-Cheol;Rho, Nayoung;Hur, On-Sook;Yoon, Hyemyeong;Lee, Myung-Chul;Oh, Sejong;Shin, Myoung-Jae;DESTA, Kebede Taye
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2019년도 추계학술대회
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    • pp.13-13
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    • 2019
  • Soybean (Glycine max L. (Merr) is commonly consumed and found in major foods including soymilk, soy sauce, tofu, and soy sprout in Korea and east Asia. In addition, it is common to cook the whole seeds with rice. Soybean is known to have ranges of health benefits including antiaging, anticancer, neuroprotective and antidiabetic taken either as supplement or dietary food. Anthocyanins and flavonoids in G. max are found to be the main contributors to such wide arrays of health benefits. Due to increasing economic values of soybean, development of specialty soybean cultivars is becoming an area of interest worldwide. In this study, 746 black soybean accessions from National Agrobiodiversity Center were characterized as part of an attempt to identify important germplasms of G. max. Seed coats of each accession were analyzed for their total anthocyanin, cyanidin 3-O-Glucoside (C-3-O-G), delphinidin 3-O-glucoside (D-3-O-G), petunidin-3-O-glucoside (Pt-3-O-G), and their whole seeds for crude protein contents. HPLC was used to determine and quantify the anthocyanin compositions while crude protein was determined using Kjeldahl method by Kjeltec auto-analyzer (Kjeltec 8400, Foss, Sweden). Accessions were grouped according to their anthocyanins and protein contents; the mean content of which were correlated to agronomic traits including maturity date, one hundred seed weight, cotyledon color and seed lust color. The results indicated that the total anthocyanin content (TAC) ranged from 273.77 to 6250.52 mg/100 g, with mean value of 1853.03 mg/100 g while the crude protein content (CPC) being between 33.43 and 47.51%, with mean value of 40.81%. The highest number of accessions (45.97%) showed TAC between 1000~1900 mg/100 g while 30.96% of accessions showed CPC between 41~43%. Among the 746 accessions considered, 11 (IT142935, 175818, 175855, 177191, 177209, 177211, 177214, 177216, 177218, 177220, 177274) of them showed TAC above 4000 mg/100 g. C-3-O-G was found to be the major contributor to TAC showing strong correlation. Accessions with green cotyledon color showed high mean TAC compared to those having yellow cotyledon color, and accessions with dull seed lust color showed high mean TAC than those having shiny seed lust color. One hundred seeds weight and maturity date showed positive correlation with all anthocyanin contents, except for Pt-3-O-G in the latter case. The overall result of the present study could be used as background for developing new black soybean cultivars and breeds with high anthocyanin and protein contents. The result depicted that many of the accessions could be used as potential parental lines.

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The Spotted Flounder (Verasper variegatus) Growth Hormone cDNA and Its Evolutionary Implications

  • Lee Jeong-Ho;Lee Sang-Jun;Kim Kyung-Kil;Kim Woo-Jin;Park Doo-Won;Park Jung-Youn
    • Fisheries and Aquatic Sciences
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    • 제6권4호
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    • pp.180-186
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    • 2003
  • The full-length cDNA encoding the pre-protein growth hormone (sfGH) from spotted flounder (Verasper variegatus) was amplified by the rapid amplification of cDNA ends (RACE) using degenerated oligonucleotide primers derived from conserved growth hormone sequences. It consists of 901 nucleotides in length, including the coding region of 609 nucleotides, 111 nucleotides of a 5' untranslated region, and 181 nucleotides of a 3' untranslated region. The conserved polyadenylation signal (AATAAA) lies 12 bases upstream from the poly (A) tail. The deduced amino acid sequence shows an open reading frame encoding a pre-protein of 203 amino acids and a putative signal peptide of 17 amino acids, suggesting that the mature hormone consists of 186 amino acids. The analyses of sfGH reveal some unique structural features. The repetitive sequences are located in the 5' untranslated region of sfGH cDNA and consist of tandem arrays of imperfect direct repeat monomers. Moreover, sfGH contains six Cys residues, as opposed to four or five in other GHs, and it is clearly distinguishable from olive flounder (Paralichthys olivaceus) GH, which lacks a region corresponding to residues 175-188 in alignment positions. It has important implications from an evolutionary standpoint, suggesting possible divergence among flatfishes.