• 제목/요약/키워드: terminal region

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Molecular Cloning, Characterization and Expression Analysis of an ILF2 Homologue from Tetraodon nigroviridis

  • Wang, Hui-Ju;Shao, Jian-Zhong;Xiang, Li-Xin;Shen, Jia
    • BMB Reports
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    • 제39권6호
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    • pp.686-695
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    • 2006
  • Interleukin-2 enhancer binding factor 2 (ILF2) was reported to regulate transcription of interleukin-2 (IL-2), a central cytokine in the regulation of T-cell responses. This property of ILF2 was well characterized in human and mammals, but little is known in bony fish. In this paper, an ILF2 homologue was cloned and well characterized from Tetraodon nigrovirid is for the further investigation of the function of ILF2 in bony fish. The full-length Tetraodon ILF2 cDNA was 1380 bp in size and contained an open reading frame (ORF) of 1164 bp that translates into a 387 amino-acid peptide with a molecular weight of 42.9 kDa, a 5' untranslated region (UTR) of 57 bp, and a 3' UTR of 159 bp containing a poly A tail. The deduced peptide of Tetraodon ILF2 shared an overall identity of 58%~93% with other known ILF2 sequences, and contained two N-glycosylation sites, two N-myristoylation sites, one RGD cell attachment sequence, six protein kinase C phosphorylation sites, one amino-terminal RGG-rich single-stranded RNA-binding domain, and a DZF zinc-finger nucleic acid binding domain, most of which were highly conserved through species compared. Constitutive expression of Tetraodon ILF2 was observed in all tissues examined, including gill, gut, head kidney, spleen, liver, brain and heart. The highest expression was detected in heart, followed by liver, head kidney and brain. Stimulation with LPS did not significantly alter the expression of Tetraodon ILF2. Gene organization analysis showed that the Tetraodon ILF2 gene have fifteen exons, one more than other known ILF2 genes in human and mouse. Genes up- and down-stream from the Tetraodon ILF2 were Rpa12, Peroxin-11b, Smad4, Snapap and Txnip homologue, which were different from that in human and mouse.

LebZIP2 induced by salt and drought stress and transient overexpression by Agrobacterium

  • Seong, Eun-Soo;Kwon, Soon -ung;Ghimire, Bimal Kumar;Yu, Chang-Yeon;Cho, Dong-Ha;Lim, Jung-Dae;Kim, Kyoung-Su;Heo, Kweon;Lim, Eun-Sang;Chung, Ill-Min;Kim, Myong-Jo;Lee, Youn-Su
    • BMB Reports
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    • 제41권10호
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    • pp.693-698
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    • 2008
  • The full-length cDNA of LebZIP2 (Lycopersicon esculentum bZIP2) encodes a protein of 164 amino acids and contains a N-terminal basic-region leucine zipper domain. Analysis of the deduced tomato LebZIP2 amino acid sequence revealed that it shares 85% sequence identity with both tobacco bZIP and pepper CcbZIP. LebZIP2 mRNA is expressed at a high level exclusively in flowers. Presently, LebZIP2 was strongly increased also following NaCl and mannitol treatments. No significant LebZIP2 expression was evident following cold treatment. Transient LebZIP2 overexpression resulted in increased NbNOA1 and NbNR transcript levels in Nicotiana benthamiana leaves. Our results indicate that LebZIP2 might play roles as an abiotic stress-signaling pathway and as a transcriptional regulator of the NbNOA1 or NbNR genes.

지능형 홈네트워크에서 네트워크 기반의 이동성 지원을 위한 P2P-SIP 구조의 설계 및 성능분석 (Design and Performance Analysis of A Novel P2P-SIP Architecture for Network-based Mobility Support in Intelligent Home Networks)

  • 김승원;정종필
    • 정보처리학회논문지:컴퓨터 및 통신 시스템
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    • 제2권7호
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    • pp.293-302
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    • 2013
  • 홈네트워크 사업자는 빠르게 확대 되는 시장 환경에 맞춰 확장성, 신뢰성, 유연성, 저비용의 구조로 홈네트워크 서비스를 제공하는 것에 대해 많은 고민을 하고 있다. 기존 클라이언트-서버 방식은 홈네트워크를 서비스하는데 있어 복잡성, 고비용 등 다양한 문제가 존재한다. 본 논문에서는 이러한 문제를 해결하기 위한 P2P-SIP(Peer to peer - Session Initiation Protocol) 구조를 제안한다. 분산된 자원의 접근을 지원하는 P2P(Peer to peer) 통신 방식의 단말에서 기존의 SIP(Session Initiation Protocol)기반 네트워크 장치들이 가지던 기능을 제공한다. 또한 단말의 이동성을 고려한 네트워크 기반의 PMIPv6(Proxy Mobile IPv6)기술을 적용하여 이동성 관리에 따른 지연시간을 최소화한다. 특히 제안하는 P2P-SIP 구조는 평면 구조에 비해 지역 내 다양한 홈네트워크 간의 뛰어난 확장성과 유지비용을 절감하는 매우 효율적인 구조임을 보여줬다.

IgE Binding Reactivity of Peptide Fragments of Bla g 4, a Major German Cockroach Allergen

  • Shin, Kwang-Hyun;Jeong, Kyoung-Yong;Hong, Chein-Soo;Yong, Tai-Soon
    • Parasites, Hosts and Diseases
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    • 제47권1호
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    • pp.31-36
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    • 2009
  • Cockroaches have been recognized as a major cause of asthma. Bla g 4 is one of the most important German cockroach allergens. The aim of this study is to investigate IgE reactivity to the recombinant Bla g 4 (rBla g 4) in the sera of allergic patients and identify linear IgE binding epitope. For protein expression, full-length Bla g 4 (EF202172) was divided into 5 overlapping peptide fragments (E1: aa 1-100, E2: aa 34-77, E3: aa 74-117, E4: aa 114-156, and E5: aa 153-182). The full-length and 5 peptide fragments of Bla g 4 was generated by PCR and over-expressed in E. coli BL21 (DE3). The IgE binding reactivities of the full-length and peptide fragments were measured by ELISA using 32 serum samples of cockroach allergy. The sera of 8 patients (25%) reacted with rBla g 4. Four sera (100%) showed IgE-binding reactivity to full-length and peptide fragment 4, and 2 sera (50%) reacted with peptide fragment 2. One (20%) serum reacted with peptide fragment 3. The results of ELISA using overlapping recombinant fragments indicated that the epitope region was located at amino acid sequences 34-73 and 78-113, and major IgE epitope of Bla g 4 was located at amino acid sequences 118-152 of C-terminal. B-cell epitope analysis of German cockroach allergen Bla g 4 could contribute to the strategic development of more specific and potentially efficacious immunotherapy.

Screening and Molecular Cloning of a Protective Antigen from the Midgut of Haemaphysalis longicornis

  • Hu, Yonghong;Zhang, Jincheng;Yang, Shujie;Wang, Hui;Zeng, Hua;Zhang, Tiantian;Liu, Jingze
    • Parasites, Hosts and Diseases
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    • 제51권3호
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    • pp.327-334
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    • 2013
  • Vaccination is considered a promising alternative for controlling tick infestations. Haemaphysalis longicornis midgut proteins separated by SDS-PAGE and transferred to polyvinylidene difluoride (PVDF) membrane were screened for protective value against bites. The western blot demonstrated the immunogenicity of 92 kDa protein (P92). The analysis of the P92 amino acid sequence by LC-MS/MS indicated that it was a H. longicornis paramyosin (Hl-Pmy). The full lenghth cDNA of Hl-Pmy was obtained by rapid amplification of cDNA ends (RACE) which consisted of 2,783 bp with a 161 bp 3' untranslated region. Sequence alignment of tick paramyosin (Pmy) showed that Hl-Pmy shared a high level of conservation among ticks. Comparison with the protective epitope sequence of other invertebrate Pmy, it was calculated that the protective epitope of Hl-Pmy was a peptide (LEEAEGSSETVVEMNKKRDTE) named LEE, which was close to the N-terminal of Hl-Pmy protein. The secondary structure analysis suggested that LEE had non-helical segments within an ${\alpha}$-helical structure. These results provide the basis for developing a vaccine against biting H. longicornis ticks.

Analysis of Symptom Determinant of Cucumber mosaic virus RNA3 via Pseudorecombinant Virus in Zucchini Squash

  • Choi, Seung-Kook;Yoon, Ju-Yeon;Choi, Jang-Kyung;Kim, Kook-Hyung;Sohn, Seong-Han
    • The Plant Pathology Journal
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    • 제23권4호
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    • pp.272-280
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    • 2007
  • Isolates of Cucumber mosaic virus (CMV) collected in Korea, were compared with their pathological features in tobacco and zucchini squash. Full-length cDNA clone of RNA3 was generated by using long-distance RT-PCR. Transcript RNA3 from the cDNA clone was inoculated onto host plants with transcripts RNA1 and RNA2 of Fny strain, generating RNA3-pseudorecombinant CMV. Timing and severity of systemic symptom was not significantly different among the pseudorecombinant CMVs in tobacco, compared with strains Fny-CMV and Pf-CMV. However, the pseudorecombinant CMVs induced two different systemic symptoms (mosaic vs. chlorotic spot) in zucchini squash. Based on symptom induction, the pseudorecombinant CMVs were categorized into two classes. The severity and timing of symptoms were correlated with viral RNA accumulations in systemic leaves of zucchini squash, suggesting that different kinetics of virus movement associated with CMV proteins are crucial for systemic infection and symptom development in zucchini squash. The analysis of movement proteins (MP) of CMV strains showed high sequence homology, but the differences of several amino acids were found in the C-terminal region between Class-I-CMV and Class-II-CMV. The analysis of coat proteins (CP) showed that the CMV isolates tested belonged to CMV subgroup I and the viruses shared overall 87-99% sequence identity in their genomes. Phylogenetic analysis of MP and CP suggested that biological properties of Korean CMV isolates have relationships associated with host species.

Production of a anti-MUC1 monoclonal antibody using a glutathione- S-transferase-MUC1 bacterial fusion protein.

  • Park, Kyu-Hwan;Shin, Chan-Young;You, Byung-Kwon;Ko, Kwang-Ho
    • 한국응용약물학회:학술대회논문집
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    • 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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    • pp.198-198
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    • 1998
  • Muc1 mucin is found in a variety of epithelial tissue and is overexpressed in several epithelial cancer. Recently it is alsol reported that primary Hamster tracheal surface epithelial(HTSE) cells express Muc1 protein and cDNA encoding HTSE muc1 protein has been cloned. Although numerous monoclonal antibodies (mAbs) to human muncins, particularly Muc1 have been produced, no such antibodies to murine Muc1 have been described. We now describe monoclonal antibody, called mAb M1CT, produced to C-terminal region of HTSE Muc1 protein by immunising mice with a glutathion-s-transferase linked fusion protein. In this study, using this antibody(mAb M1CT) we investigated the effect of RA on the expression of Muc1 in HTSE cells. Retinoic acid(RA) plays an essential role in maintaining normal differentiation of tracheal epithelial cells. With RA-deficiency tracheocytes undergo squamous metaplasia, an abnormal differentiation that can be reversed by RA. We had primary culture of HTSE cells under different concentrations of RA. Culture was maintained until the direction of differentiation was determined. Then Western blot analysis with mAb M1CT was performed with the cell lysates from the culture. The expression of Muc1 protein was decreased in dose-dependent manner as the concentration of retinoic acid was decreased. Our result indicates that the expression of Muc1 protein is coordinately regulated with airway mucous cell differentiation by RA pathway. And the antibody, mAb M1CT, produced in this study should provide useful tool to study the expression of Muc1 mucin in differentiation process or disease.

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Bi-Directional Half-Duplex Relaying Protocols

  • Kim, Sang-Joon;Devroye, Natasha;Tarokh, Vahid
    • Journal of Communications and Networks
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    • 제11권5호
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    • pp.433-444
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    • 2009
  • The bi-directional relay channel is the natural extension of a three-terminal relay channel where node a transmits to node b with the help of a relay r to allow for two-way communication between nodes a and b. That is, in a bi-directional relay channel, a and b wish to exchange independent messages over a shared channel with the help of a relay r. The rates at which this communication may reliably take place depend on the assumptions made on the relay processing abilities. We overview information theoretic limits of the bi-directional relay channel under a variety of conditions, before focusing on half-duplex nodes in which communication takes place in a number of temporal phases (resulting in protocols), and nodes may forward messages in four manners. The relay-forwarding considered are: Amplify and forward (AF), decode and forward (DF), compress and forward (CF), and mixed forward. The last scheme is a combination of CF in one direction and DF in the other. We derive inner and outer bounds to the capacity region of the bi-directional relay channel for three temporal protocols under these four relaying schemes. The first protocol is a two phase protocol where a and b simultaneously transmit during the first phase and the relay r alone transmits during the second. The second protocol considers sequential transmissions from a and b followed by a transmission from the relay while the third protocol is a hybrid of the first two protocols and has four phases. We provide a comprehensive treatment of protocols in Gaussian noise, obtaining their respective achievable rate regions, outer bounds, and their relative performance under different SNR and relay geometries.

Identification and Functional Analysis of Mating Type Loci in the Pleurotus eryngii

  • Ryu, Jae San;Kim, Min-Keun;Park, Bokyung;Ali, Asjad;Joung, Wan-Kyu
    • 한국균학회소식:학술대회논문집
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    • 한국균학회 2015년도 추계학술대회 및 정기총회
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    • pp.35-35
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    • 2015
  • Pleurotus eryngii has recently become a major cultivated mushroom; it uses tetrapolar heterothallism as a part of its reproductive process. Sexual development progresses only when the A and B mating types are compatible. Such mating incompatibility occasionally limits the efficiency of breeding programs in which crossing within loci-shared strains or backcrossing strategies are employed. Therefore, understanding the mating system in edible mushroom fungi will help provide a short cut in the development of new strains. We isolated and identified pheromone and receptor genes in the B3 locus of P. eryngii and performed a functional analysis of the genes in the mating process by transformation. A genomic DNA library was constructed to map the entire mating-type locus. The B3 locus was found to contain four pheromone precursor genes and four receptor genes. Remarkably, receptor PESTE3.3.1 has just 34 amino acid residues in its C-terminal cytoplasmic region; therefore, it seems likely to be a receptor-like gene. Real-time quantitative RT-PCR (real-time qRT-PCR) revealed that most pheromone and receptor genes showed significantly higher expression in monokaryotic cells than dikaryotic cells. The pheromone genes PEphb3.1 and PEphb3.3 and the receptor gene PESTE3.3.1 were transformed into P5 (A3B4). The transformants were mated with a tester strain (A4B4), and the progeny showed clamp connections and a normal fruiting body, which indicates the proposed role of these genes in mating and fruiting processes. This result also confirms that PESTE3.3.1 is a receptor gene. In this study, we identified pheromone and receptor genes in the B3 locus of P. eryngii and found that some of those genes appear to play a role in the mating and fruiting processes. These results might help elucidate the mechanism of fruiting differentiation and improve breeding efficiency.

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CND41, a DNA-binding protein in chloroplast nucleoid, and its function

  • Sato, Fumihiko;Murakami, Shinya;Chatani, Hiroshi;Nakano, Takeshi
    • 한국식물학회:학술대회논문집
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    • 한국식물학회 1999년도 제13회 식물생명공학심포지움 New Approaches to Understand Gene Function in Plants and Application to Plant Biotechnology
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    • pp.51-56
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    • 1999
  • Plastids, which are organelles unique to plant cells, bear their own genome that is organized into DNA-protein complexes (nucleoids). Regulation of gene expression in the plastid has been extensively investigated because this organelle plays an important role in photosynthesis. Few attempts, however, have been made to characterize the regulation of plastid gene expression at the chromosomal structure, using plastid nucleoids. In this report, we summarize the recent progress in the characterization of DNA-binding proteins in plastids, with special emphasis on CND41, a DNA binding protein, which we recently identified in the choloroplast nucleoids from photomixotrophically cultured tobacco cells. CND41 is a protein of 502 amino acids which consisted of a transit peptide of 120 amino acids and a mature protein of 382 amino acids. The N-terminal of the 'mature' protein has lysine-rich region which is essential for DNA-binding. CNA41 also showed significant identities to some aspartyl proteases. Protease activity of purified CND41 has been recently confirmed and characterized. On the other hand, characterization of accumulation of CND41 both in wild type and transgenic tobacco with reduced amount of CND41 suggests that CND41 is a negative regulator in chloroplast gene expression. Further investigation indicated that gene expression of CND41 is cell-specifically and developmentally regulated as well as sugar-induced expression. The reduction of CND41 expression in transgenic tobacco also brought the stunted plant growth due to the reduced cell length in stem. GA3 treatment on apical meristem reversed the dwarf phenotype in the transformants. Effects of CND41 expression on GA biosynthesis will be discussed.

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