• 제목/요약/키워드: Brain cDNA library

검색결과 58건 처리시간 0.025초

Direct and functional interaction between dopamine D2 receptor and ALY

  • Yang, Ji-Hye;Cheong, Da-Woon;Seo, Hyung-Ju;Kim, Moon-Soo;Kim, Kyeong-Man
    • 대한약학회:학술대회논문집
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    • 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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    • pp.270.1-270.1
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    • 2002
  • The signaling pathway of D2 dopamine receptor was studied using yeaslt two-hybrid system.. The 3rd cytoplasmic loop of rat D2 dopamine receptor was used to screen the cDNA library of mouse brain. and ALY was found to interact with it. The interaction in the yeast was observed only with the 3rd cytoplasmic loop of D2 dopamine receptor but not with that of D3 or D4 dopamine receptor. The interaction between two proteins was also confirmed by GST pull-down assay. (omitted)

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Calmodulin of Olive Flounder Paralichthys olivaceus : Cloning and Expression Analysis

  • Hong, Gyeong-Eun;Kong, Hee Jeong;Nam, Bo-Hye;Kim, Young-Ok;Kim, Woo-Jin;Lee, Sang-Jun;Choi, Tae-Jin
    • 한국해양바이오학회지
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    • 제2권4호
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    • pp.234-237
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    • 2007
  • Calmodulin은 $Ca^{2+}$ 결합단백질로서 생체 내에서 $Ca^{2+}$ 의존적 기작을 통하여 다양한 생물학적 기능에 관여한다. 본 연구에서는 넙치 Paralichthys olivaceus의 cDNA library로부터 Calmodulin cDNA를 분리 동정하였다. 염기 서열 및 아미노산 서열을 분석한 결과, 넙치 Calmodulin cDNA는 782개의 nucleotides로 구성되어 있고, 4개의 잘 보존된 $Ca^{2+}$결합 motifs (EF-I, EF-II, EF-III, EF-IV)를 가지는 149개의 아미노산 잔기를 전사할 수 있는 open reading frame을 포함한다. 또한 번역된 아미노산 서열은 인간, 쥐, zebrafish, 개구리의 Calmodulin 아미노산 서열과 100% 동일성을 보이며 보라성게, 침팬지의 Calmodulin 아미노산 서열과 각각 97, 99%의 동일성을 보인다. 넙치 Calmodulin 전사체는 뇌와 장 조직에서 많은 양이 발현되었고, 신장, 아가미, 눈, 근육, 피부, 지느러미에서도 발현이 관찰되었다. 또한 넙치 Calmodulin 전사체는 수정 후 7일째의 발생 초기 단계 시료에서도 발현되어 수정 후 34일째까지 그 발현이 서서히 증가하였다. 이상의 결과들로부터 넙치 Calmodulin은 넙치의 발생 초기 단계에서 필요한 단백질로 생각되며 아마 항상성 유지에 중요한 역할을 할 것으로 예상된다.

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Construction of EST Database for Comparative Gene Studies of Acanthamoeba

  • Moon, Eun-Kyung;Kim, Joung-Ok;Xuan, Ying-Hua;Yun, Young-Sun;Kang, Se-Won;Lee, Yong-Seok;Ahn, Tae-In;Hong, Yeon-Chul;Chung, Dong-Il;Kong, Hyun-Hee
    • Parasites, Hosts and Diseases
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    • 제47권2호
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    • pp.103-107
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    • 2009
  • The genus Acanthamoeba can cause severe infections such as granulomatous amebic encephalitis and amebic keratitis in humans. However, little genomic information of Acanthamoeba has been reported. Here, we constructed Acanthamoeba expressed sequence tags (EST) database (Acanthamoeba EST DB) derived from our 4 kinds of Acanthamoeba cDNA library. The Acanthamoeba EST DB contains 3,897 EST generated from amebae under various conditions of long term in vitro culture, mouse brain passage, or encystation, and downloaded data of Acanthamoeba from National Center for Biotechnology Information (NCBI) and Taxonomically Broad EST Database (TBestDB). The almost reported eDNA/genomic sequences of Acanthamoeba provide stand alone BLAST system with nucleotide (BLAST NT) and amino acid (BLAST AA) sequence database. In BLAST results, each gene links for the significant information including sequence data, gene orthology annotations, relevant references, and a BlastX result. This is the first attempt for construction of Acanthamoeba database with genes expressed in diverse conditions. These data were integrated into a database (http://www. amoeba.or.kr).

ZNF435, a Novel Human SCAN-containing Zinc Finger Protein, Inhibits AP-1-mediated Transcriptional Activation

  • Gu, Xing;Zheng, Mei;Fei, Xiangwei;Yang, Zhenxing;Li, Fan;Ji, Chaoneng;Xie, Yi;Mao, Yumin
    • Molecules and Cells
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    • 제23권3호
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    • pp.316-322
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    • 2007
  • Zinc finger transcription factor genes are a significant fraction of the genes in the vertebrate genome. Here we report the isolation and characterization of a human zinc finger-containing gene, ZNF435, from a fetal brain cDNA library. ZNF435 cDNA is 1290 base pairs in length and contains an open reading frame encoding 349 amino acids with four C2H2-type zinc fingers at its carboxyl terminus and a SCAN motif at its amino terminus. RT-PCR results showed that ZNF435 was expressed in all tested tissues. A ZNF435-GFP fusion protein was located in the nucleus and the four zinc fingers acted as nuclear localization signals (NLSs). ZNF435 was found to be capable of homo-association, and this effect was independent of its zinc fingers. Furthermore, ZNF435 proved to be a transcription repressor as its overexpression in AD293 cells inhibited the transcriptional activities of AP-1.

인회석 박막 피복 도관과 Brain-derived neurotrophic factor(BDNF) 유전자 이입 슈반세포를 이용한 백서 좌골신경 재생에 관한 연구 (SCIATIC NERVE REGENERATION USING CALCIUM PHOSPHATE COATED CONDUIT AND BRAIN-DERIVED NEUROTROPHIC FACTOR GENE-TRANSFECTED SCHWANN CELL IN RAT)

  • 최원재;안강민;황순정;정필훈;김명진;김남열;유상배;장정원;김현만;김중수;김윤희;김성민;이종호
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제31권3호
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    • pp.199-218
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    • 2005
  • Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.

Surface expression of TTYH2 is attenuated by direct interaction with β-COP

  • Ryu, Jiwon;Kim, Dong-Gyu;Lee, Young-Sun;Bae, Yeonju;Kim, Ajung;Park, Nammi;Hwang, Eun Mi;Park, Jae-Yong
    • BMB Reports
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    • 제52권7호
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    • pp.445-450
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    • 2019
  • TTYH2 is a calcium-activated, inwardly rectifying anion channel that has been shown to be related to renal cancer and colon cancer. Based on the topological prediction, TTYH2 protein has five transmembrane domains with the extracellular N-terminus and the cytoplasmic C-terminus. In the present study, we identified a vesicle transport protein, ${\beta}$-COP, as a novel specific binding partner of TTYH2 by yeast two-hybrid screening using a human brain cDNA library with the C-terminal region of TTYH2 (TTYH2-C) as a bait. Using in vitro and in vivo binding assays, we confirmed the protein-protein interactions between TTYH2 and ${\beta}$-COP. We also found that the surface expression and activity of TTYH2 were decreased by co-expression with ${\beta}$-COP in the heterologous expression system. In addition, ${\beta}$-COP associated with TTYH2 in a native condition at a human colon cancer cell line, LoVo cells. The over-expression of ${\beta}$-COP in the LoVo cells led to a dramatic decrease in the surface expression and activity of endogenous TTYH2. Collectively, these data suggested that ${\beta}$-COP plays a critical role in the trafficking of the TTYH2 channel to the plasma membrane.