• 제목/요약/키워드: Annealing control primer

검색결과 47건 처리시간 0.035초

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

  • 이상범;진형주
    • 한국어류학회지
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    • 제24권2호
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    • pp.118-124
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    • 2012
  • 이전 연구에서 넙치유래 재조합 마이오스타틴 프로도메인을 무지개송어에 한달간 침지법을 통하여 처리한 결과 대조군에 비하여 무게가 최대 약 42% 증가되었다. 따라서 본 연구는 재조합 마이오스타틴 프로도메인을 침지법에 의해 처리된 무지개송어와 대조군의 근육으로부터 발현되는 cDNA를 제작하여 마이오스타틴 프로도메인에 의해서 유도된 특정유전자를 선발하기 위하여 ACP (annealing control primer)를 이용한 DDRT법을 통하여 분석하였다. 총 20가지의 ACP를 이용한 결과 2개의 특정 유전자를 분석하였으며, NCBI BLAST 분석결과 Cytochrome P450 mono oxygenase와 Profilin으로 판명되었다. 이 중 Cytochrome P450 mono oxygenase는 대조군보다 발현량이 증가하였으며, Profilin는 대조군에 비해서 발현량이 감소하였다. 이러한 결과를 재확인하기 위하여 두 유전자의 primer를 각각 제작하여 semi-quantitative RT-PCR를 시행한 결과 DDRT법에 의한 분석과 동일하였다. 본 결과는 어류의 성장에서 마이오스타틴 프로도메인의 기능 및 메카니즘에 대한 연구에 유용한 자료가 될 것으로 사료된다.

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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    • 제14권1호
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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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    • 제37권3호
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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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    • 제16권3호
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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
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2004년도 춘계학술발표대회
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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
    • 한국초지조사료학회지
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    • 제36권3호
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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.

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

  • 이진성;홍수영;이기화
    • 생명과학회지
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    • 제20권6호
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    • pp.929-933
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    • 2010
  • 본 연구는 곤충으로부터 새로운 항 진균 단백질을 발굴하기 하기 위한 목적으로 누에를 대상으로 Aspergillus niger의 감염을 유도하였을 때 발현되는 유전자의 특성을 분석한 것이다. Annealing control primer 법에 기초한 GeneFishing Kit를 사용하여 A. niger를 약 $6{\times}10^8$ colony per unit로 5령기 누에 유충의 체강에 감염시킨 후, 6시간 경과한 다음에 유도 발현되는 유전자(differentially expressed genes, DEGs)를 분석 한 결과, 10개의 유도 발현되는 유전자를 분리하였고 RT-PCR을 통해서 lysozyme, enbocin 그리고 한 개의 기능이 알려지지 않는 유전자등 3개의 유전자가 A. niger의 감염에 의해서 유의하게 과 발현된다는 것을 검증하였다. 일반적으로 그람 음성 및 양성 세균의 감염에 의해 유도된다고 알려진 enbocin 유전자가 A. niger의 감염에서도 과 발현이 유도되는 본 연구의 결과는 앞으로 enbocin 유전자의 항 진균 활성 연구에 중요한 기초 자료로 활용될 수 있을 것이다.