• Title/Summary/Keyword: Gfp

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Encapsulation of IP Traffic through GFP in OTN Transmission Network (OTN 광전송망에서 GFP를 통한 IP 트래픽의 인캡슐레이션)

  • Lee, Chang-Ki;Yang, Choong-Reol
    • The KIPS Transactions:PartC
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    • v.15C no.6
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    • pp.567-572
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    • 2008
  • It is necessary to study about the scheme to accept IP traffic effectively in OTN, according to expect that IP traffic data will be increase constantly. In this paper, we studied the encapsulation method of IP traffic through GFP in OTN transmission network. Therefore we knew the IP/GFP/OTN method is more efficient than existing methods from structure, overhead rates and possibility of grow, and showed the functional block of high level about this method. Also we showed the implementation scheme of this method by processing the functional simulation to make use of VHDL programming.

Optimization of Gene Transfection Using Fluorescence-Activated Cell Sorter(FACS) Analysis of Green Fluorescent Protein(GFP) (Green Fluorescent Protein(GFP)의 Fluorescence-Activated Cell Sorter(FACS) 분석을 통한 유전자 이입의 최적화)

  • 김태경;박민태;이균민
    • KSBB Journal
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    • v.14 no.3
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    • pp.377-379
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    • 1999
  • In order to improve the transfection efficiency of CHO/dhfr- cells using cationic lipid, optimal concentrations of the cationic lipid($LipofectAmine^{TM}$) and DNA(pEGFP-C1) need to be determined. The use of green fluorescent protein(GFP) gene as a reporter gene facilitated the quantification of transfection efficiency. The green fluorescence intensity of each cell transfected at various lipid-DNA concentrations was measured using fluorescence-activated cell sorter(FACS) analysis. A combination of $2.0{\mu}L$ cationic lipid and 0.4{$\mu}g$ DNA in a well resulted in the highest trasfection efficiency. Taken together, the method using FACS analysis of GFP is simple and fast, facilitating the optimization of transfection.

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The Use of a Tobacco mosaic virus-Based Expression Vector System in Chrysanthemum

  • Park, Minju;Baek, Eseul;Yoon, Ju-Yeon;Palukaitis, Peter
    • The Plant Pathology Journal
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    • v.33 no.4
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    • pp.429-433
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    • 2017
  • Chrysanthemums (Chrysanthemum morifolium) are susceptible to tobacco mosaic virus (TMV). TMV-based expression vectors have been used in high-throughput experiments for production of foreign protein in plants and also expressing green fluorescent protein (GFP) to allow visualization of TMV movement. Here, we used TMV expressing the GFP to examine the infection of chrysanthemum by a TMV-based expression vector. Viral replication, movement and GFP expression by TMV-GFP were verified in upper leaves of chrysanthemums up to 73 days post inoculation (dpi) by RT-PCR. Neither wild-type TMV nor TMV-GFP induced symptoms. GFP fluorescence was seen in the larger veins of the inoculated leaf, in the stem above the inoculation site and in petioles of upper leaves, although there was no consistent detection of GFP fluorescence in the lamina of upper leaves under UV. Thus, a TMV-based expression vector can infect chrysanthemum and can be used for the in vivo study of gene functions.

GFP expression in the microspore-derived early embryo through co-culturing with Agrobacterium (Agrobacterium 공동배양을 이용한 고추 소포자 유래 초기 배의 GFP 발현)

  • Jung, Min;In, Dong-Su;Kim, Bong-Kyu;Jang, In-Chang;Park, Eun-Joon;Kim, Moon-Za;Harn, Chee-Hark
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.109-114
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    • 2008
  • The aim of this research is to establish the conditions for Agrobacterium-mediated genetic transformation using microspore. The embryo induction from the microspore was examined under several Kanamycin concentration in media, and the induction rate decreased about 4, 8, 10 times when the Kanamycin concentration increased 10, 50, 100 mg/L, respectively. This indicates that the transformation rate would be much lower if the Kanamycin was used for selection marker. In order to apply the GFP gene as a reporter gene for Agrobacterium-mediated genetic transformation, GFP expression from the microspore-mediated embryos was observed using GFP filter under microscope. The GFP expression occurred when the microspore cultured toward the embryo development for 12, 24 and 48 days. The microspore formed a cluster by microspore division from 12 days culture and continuously became a bigger mass. We obtained a total of 8 GFP-expressing embryos suggesting that the transformation of microspore occurred. However, those young embryos were not fully developed. Further study pertinent to culture conditions is required to fulfill the Agrobacterium-mediated genetic transformation using microspore.

Analysis of the Foreign Gene Transmission in the GFP Transgenic Chickens (형질전환 닭에서 GFP 유전자 전이 연구)

  • Jang, Ye-Jin;Ji, Mi-Ran;Jeon, Mi-Hyang;Kim, Jeom-Sun;Kim, Kyung-Woon;Han, Deug-Woo;Chung, Hak-Jae;Yang, Byoung-Chul;Yoo, Jae-Gyu;Park, Jin-Ki;Kim, Te-Oan;Byun, Sung-June
    • Korean Journal of Poultry Science
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    • v.39 no.3
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    • pp.241-244
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    • 2012
  • This study was performed to analyze the generational transmission and the expression of the foreign gene in the GFP transgenic chickens. The transmission rate and the expression of the GFP gene was investigated from the GFP transgenic rooster (G2) as the first founder to the ninth (G8). Analysis of GFP expression in hatched chickens was used the UV lamp. When GFP was expressed in the wings, bill and legs of a chick, the bird only was selected as a transgenic chick. The average transmission rate of the overall transgenic was 38~58%. These results showed that the transmission of the GFP gene in the transgenic chickens in accordance with the laws of Mendel's continues to the next generation without gene silencing.

Cell growth and GFP expression in E. coli BL21 and W3110 under coexpression of Vitreoscilla hemogobin

  • Gang, Dong-Gyun;Kim, Yeon-Gyu;Cha, Hyeong-Jun
    • 한국생물공학회:학술대회논문집
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    • 2001.11a
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    • pp.754-757
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    • 2001
  • Expression of the vhb gene encoding bacterial hemoglobin (VHb) from Vitreoscilla has been used to improve recombinant cell growth and enhance product formation under microaerobic conditions because of its ability to enhance oxygen use. We coexpressed GFP and VHb in Escherichia coli BL21 and W3110, and compared with GFP control which was not expressed VHb. We used nar oxygen-dependent inducible promoter for VHb expression. The GFP amounts in E. coli expressed VHb was about five fold higher than in the control Fluorescence intensity was increased about two fold.

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GFP Gene Transfected Cell 과 Non Tranfsfected Cell 의 핵이식후 발달

  • 양병철;임기순;성환후;임석기;이상기;오현주;이연근;박진기;장원경
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.34-34
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    • 2002
  • 핵이식 방법은 형질전환 동물을 생산하기 위한 여러 가지 방법 중 최근에 많이 이용되고 있다. 본 실험은 형광단백질 유전자 (green fluorescence protein, GFP)가 도입된 태아섬유아세포를 이용 핵이식을 하여 형질전환 수정란의 생산효율을 검토하기 위하여 실시하였다. GFP 유전자는 임신 45-55 일령의 태아섬유아세포 (KbFF3)에 electroporation방법으로 transfection을 실시하였다. (중략)

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Regulation of GFP Expression Using the Tetracycline Inducible Retroviral Vector System (Tetracycline Inducible Retrovirus Vector System에 의한 GFP 유전자의 발현 조절)

  • Koo Bon Chul;Kwon Mo Sun;Kim Teoan
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.57-62
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    • 2005
  • One of the critical problems to be solved in transgenic animal production is uncontrollable constitutive expression of foreign genes, which usually results in serious physiological disturbances in the transgenic animal. To circumvent this problem, we constructed and tested two retrovirus vectors designed to express the GFP(green fluorescent protein) gene under the control of the tetracycline-inducible promoters. To maximize the GFP gene expression at turn-on state, WPRE(woodchuck hepatitis virus posttranscriptional regulatory element) sequence was introduced into the retrovirus vectors at downstream region of either the GFP gene or the sequence encoding rtTA(reverse tetracycline-controlled transactivator). Transformed cells were cultured in the medium supplemented with or without doxycycline(tetracycline derivative) for 48 hours, and induction efficiency was measured by comparing the GFP gene expression level using fluorometry and western blotting. Higher GFP expression was observed from the vector carrying the WPRE sequence at 3' side of the GFP gene, while tighter expression control(up to 20 fold) was obtained from the vector in which the WPRE sequence was placed at 3' side of rtTA sequence. The resulting tetracycline inducible vector system may be used in transgenic animal production and gene therapy.

Identification of C4orf32 as a Novel Type I Endoplasmic Reticulum Resident Membrane Protein (Type I 소포체 목표화 막단백질에 속하는 새로운 C4orf32 막단백질의 동정)

  • Lee, Seung-Hwan;Park, Sang-Won;Lee, Jin-A;Jang, Deok-Jin
    • Journal of Life Science
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    • v.29 no.9
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    • pp.949-954
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    • 2019
  • Membrane topology is a key characteristic of membrane proteins. We previously reported the cloning of the chromosome 4 open-reading frame 32 (C4orf32) gene as a potential membrane protein; however, the cellular localization and membrane topology of C4orf32 was as yet unknown. In this study, we found that green fluorescent protein (GFP) fused to the C-terminus of C4orf32 (C4orf32-GFP) was localized to the endoplasmic reticulum (ER). We applied three tools to identify determinants of C4orf32 topology: protease protection, fluorescence protease protection (FPP), and an inducible system using the ternary complex between FK506 binding protein 12 (FKBP), rapamycin, and the rapamycin-binding domain of mTOR (FRB) (the FRB-rapamycin-FKBP system). Using protease protection and FPP assays, we found that the GFP tag in C4orf32-GFP was localized to the cytoplasmic surface of the ER membrane of HeLa cells. Protease protection and FPP assays are useful and complimentary tools for identifying the topology of GFP fusion membrane proteins. The FRB-rapamycin-FKBP system was also used to study the topology of C4orf32. In the absence of rapamycin, a monomeric red fluorescent protein-FKBP fusion (mRFP-FKBP) and C4orf32-GFP-FRB were localized to the cytoplasm and the ER membrane, respectively. However, in the presence of rapamycin, the mRFP-FKBP was shifted from the cytoplasm to the ER and colocalized with the C4orf32-GFP-FRB. These results indicate that the FRB moiety is facing the cytoplasmic surface of ER membrane. Overall, our results clearly suggest that C4orf32 belongs to the family of type I ER resident membrane proteins.

Expression Comparison of the GFP Gene under the Controls of Several Internal Promoters in the Retrovirus Vectors with or without WPRE Sequence (여러 표적세포에서 Retrovirus Vector의 내부 Promoter의 종류와 WPRE의 유무에 따른 GFP 유전자의 발현 효율성 비교)

  • Kim Y. H.;Koo B. C.;Kwon M. S.;Kim T. O.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.191-196
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    • 2004
  • In this study, to construct more effective retrovirus system, we compared four internal promoters (RSV: Rous sarcoma virus, UbC: Ubiquitin C, β-actin, CMV: human cytomegalovirus) in the retrovirus vector by measuring GFP (green fluorescent protein) expression. The effect of WPRE (woodchuck hepatitis virus posttrans-criptional regulatory element) sequence on transgene expression was also investigated. Experiments were conducted with cells derived from three different species (human, pig and chicken) and evaluated the activity of each promoter and the effect of WPRE sequence by fluorometry and Western blotting. In all cells tested, RSV and CMV promoters were superior to UbC and β-actin promoters, and RSV promoter was the best one in chicken cells. The boosting effect of WPRE sequence on GFP expression was evident in human and porcine cells but not in chicken cells.