• Title/Summary/Keyword: Annealing control primer

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Identification of Differentially Expressed Genes in Improved Rainbow Trout Growth by Treatment with a Fish Myostatin Prodomain Using the Annealing Control Primer System (Annealing control primer system을 이용한 어류 재조합 myostatin prodomain 단백질에 의해 성장이 증가된 무지개송어의 특이적 발현 유전자 탐색)

  • Lee, Sang-Beum;Jin, Hyung-Joo
    • Korean Journal of Ichthyology
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    • v.24 no.2
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    • pp.118-124
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    • 2012
  • The present study was conducted to investigate different gene expression profile between treated poMSTNpro and non-treated in rainbow trout and to identify those genes that are specifically or predominantly expressed in treated poMSTNpro by employing annealing control primer (ACP)-based GeneFishing polymerase chain reaction (PCR). We isolated total RNAs in muscle tissues from the treated poMSTNpro fish by immersion bath technique with fish myostatin prodomain (Paralichthys olivaceus, poMSTNpro) for one month and the other was non-treated poMSTNpro, and synthesized cDNA using annealing control primers (ACP, Seegene, Korea). Using 20 different ACPs for PCR, were cloned sequenced, and analyzed identities of 2 differentially expressed genes (DEGs). According to BLAST analysis, sequences of 2 clones significantly matched database entries and confirmed by semi-quantitative RT-PCR. The functional roles of one up-regulated gene, cytochrome P450 mono-oxygenases 2K1v2 (CYP2K1v2), and one down-regulated gene was Profilin-1 were identified. We identified distinctive gene expression profiles in improved rainbow trout growth by treatment with a fish myostatin prodomain using ACP-based GeneFishing.

Identification of differentially expressed genes in the developmental stages from olive flounder Paralichthys olivaceus using an annealing control primer system

  • Kim, Young-Ok;Park, Eun-Mi;Nam, Bo-Hye;Kong, Hee-Jeong;Kim, Woo-Jin;Noh, Jae-Koo;Lee, Sang-Jun;Kim, Kyung-Kil
    • Animal cells and systems
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    • v.14 no.1
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    • pp.25-30
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    • 2010
  • We employed a new and improved differential display reverse transcription-polymerase chain reaction (DDRT-PCR) method, which involves annealing control primers (ACPs), to identify the genes that are specifically or prominently expressed in olive flounder (Paralichthys olivaceus) juveniles (35 days post-hatch; dph) compared to larval-stage (dph 21) flounder. Using 60 ACPs, we identified eight differentially expressed genes (DEGs) and basic local alignment search tool (BLAST) searches revealed eight known genes. Gene expression levels were confirmed by RT-PCR. Phosphoglucose isomerase (PGI) was highly expressed at 21 dph, while nephrosin, myosin light chain (MLC), myosin heavy chain (MHC), carboxypeptidase A, chymotrypsin B, fish-egg protein, and matrix protein were expressed at 35 dph. PGI, MLC, and MHC expression was further analyzed by RT-PCR. The differentially expressed genes identified in this study may provide insights into the molecular basis of development in olive flounder.

Identification of differentially expressed genes using an annealing control primer system in periodontitis

  • Na, Hee-Sam;Kim, Ji-S.;Chung, Jin
    • International Journal of Oral Biology
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    • v.37 no.3
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    • pp.109-114
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    • 2012
  • In the gingival tissues of patients with periodontitis, inflammatory responses are mediated by a wide variety of genes. In this study, we screened for differentially expressed genes (DEGs) in periodontitis compared with normal tissue using an annealing control primer (ACP) system. By ACP RT-PCR analysis, we obtained about 160 amplicons, 8 of which were found to be differentially expressed. DEGs in patients with periodontitis were thus successfully and reliably identified by the ACP-based RT PCR technique. The DEGs identified in the screen may also enhance our understanding of the pathogenesis of periodontitis.

Identification of Differentially Expressed Genes by Proto-oncogene Protein DEK using Annealing Control Primers

  • Kim, Dong-Wook;Lee, Jae-Hwi;Seo, Sang-Beom
    • Biomolecules & Therapeutics
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    • v.16 no.3
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    • pp.184-189
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    • 2008
  • The proto-oncogene protein DEK has been implicated in various human disease including cancer. We have shown that DEK induces caspase-dependent apoptosis in Drosophila by regulating histone acetylation. Reverse transcription-polymerase chain reaction (RT-PCR) method based on annealing control primers was used to screen and identify differentially expressed genes (DEGs) in DEK overexpressed HeLa cells. Among the genes identified, clusterin and fibrillarin have major role in apoptosis pathway regulation. TFIIIC and RPS24 are implicated in HAT mediated transcriptional initiation and cololectal cancer, respectively. To further analyze DEK's role in apoptosis, multiplex PCR was performed. Caspase-3, -7, and -10 and proapoptotic gene bid were newly identified as possible target genes regulated by DEK expression.

Identification of Differentially Regulated Genes in Bovine Blastocysts using an Annealing Control Primer System

  • Park, Sae-Young;Hwang, Kyu-Chan;Cui, Xiang-Shun;Shin, Mi-Ra;Kim, Eun-Young;Lee, Won-Don;Kim, Nam-Hyung;Park, Sepill;Lim, Jin-Ho
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.229-229
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    • 2004
  • The identification of embryo-specific genes would provide insights into early embryonic development. However, the current methods employed to identify the genes that are expressed at a specific developmental stage are labor intensive and suffer from high rates of false positives. Here we employed a new and accurate reverse transcription-polymerase chain reaction (RT-PCR) technology that involves annealing control primers (ACPs) to identify the genes that are specifically or prominently expressed in bovine early blastocysts and hatched blastocysts produced in vitro. (omitted)

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An Annealing Control Primer (ACP) System Used for the Isolation and Identification of Copper-Induced Genes in Alfalfa Leaves

  • Lee, Ki-Won;Lee, Sang-Hoon;Kim, Ki-Yong;Ji, Hee Chung;Park, Hyung Soo;Hwang, Tae Young;Choi, Gi Jun;Rahman, Md. Atikur
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.36 no.3
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    • pp.237-242
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    • 2016
  • Copper (Cu) is a necessary microelement for plants. However, high concentrations of Cu are toxic to plants that change the regulation of several stress-induced proteins. In this study, an annealing control primer (ACP) based approach was used to identify differentially expressed Cu-induced genes in alfalfa leaves. Two-week-old alfalfa plants (Medicago sativa L.) were exposed to Cu for 6 h. Total RNAs were isolated from treated and control leaves followed by ACP-based PCR technique. Using GeneFishing ACPs, we obtained several genes those expression levels were induced by Cu. Finally, we identified several genes including UDP-glucuronic acid decarboxylase, transmembrane protein, small heat shock protein, C-type cytochrome biogenesis protein, mitochondrial 2-oxoglutarate, and trans-2,3-enoyl-CoA reductase in alfalfa leaves. These identified genes have putative functions in cellular processes such as cell wall structural rearrangements, transduction, stress tolerance, heme transport, carbon and nitrogen assimilation, and lipid biosynthesis. Response of Cu-induced genes and their identification in alfalfa would be useful for molecular breeding to improve alfalfa with tolerance to heavy metals.

Cloning and Identification of Differentially Expressed Genes Induced by Fungal Infection from Silkworm, Bombyx mori (누에에서 곰팡이(Aspergillus niger) 감염에 의해 유도 발현되는 유전자의 클로닝과 동정)

  • Lee, Jin-Sung;Hong, Su-Young;Lee, Ki-Hwa
    • Journal of Life Science
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    • v.20 no.6
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    • pp.929-933
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    • 2010
  • We tried to identify differentially expressed genes (DEGs) from a silkworm, Bombyx mori, involved in fungal (Aspergillus niger) infection. A total RNA purified from fungal-induced and normal B. mori ($5^{th}$ instar larvae) was used for the cDNA synthesis. Differentially expressed genes were screened by annealing control primer (ACP)-based PCR technique. Comparing the gene expression profiles between fungal infection and control silkworm, we detected 10 genes that were differentially expressed in fungal induction and performed molecular cloning and nucleotide sequencing of the 10 genes. We confirmed the expression patterns of 3 DEGs by RT-PCR. The 3 DEGs over-expressed in fungal infection were identified as lysozyme, enbocin and an unknown gene. They were first identified to be genes induced by fungal infection. Although the detailed functions of 3 genes and their products remain to be determined, the genes will provide insight into the molecular mechanisms of insect-immune systems induced by fungal infection.