• Title/Summary/Keyword: L-C Library

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Construction of a full-length cDNA library from Typha laxmanni Lepech. and T. angustifolia L. from an EST dataset

  • Im, Subin;Kim, Ho-Il;Kim, Dasom;Oh, Sang Heon;Kim, Yoon-Young;Ku, Ja Hyeong;Lim, Yong Pyo
    • Korean Journal of Agricultural Science
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    • v.45 no.4
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    • pp.583-590
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    • 2018
  • Genus Typha L. (Typhaceae; Cattail in common) is one of the hydrophytic plants found in semi-aquatic regions. About nine to 18 species of the genus exist all over the world. In Korea, the most commonly found cattail species are T. laxmanni and T. angustifolia. The aim of this study was to prepare a cDNA library and sequences and analyze expressed sequence tags (ESTs) from these species, T. laxmanni and T. angustifolia. In the case of T. laxmanni, we observed that 715 out of 742 ESTs had high quality sequences, whereas the remaining 27 ESTs were low quality sequences. In this study, we identified 77 contigs, 393 unassembled clones and 65.7% singletons. Furthermore, in the case of T. angustifolia, we recorded 992 high quality EST sequences, and by excluding 28 low quality sequences from among them, we retrieved 120 contigs, 348 unassembled clones and 48.9% singletons. The basic local alignment search tool (BLAST) and Kyoto encyclopedia of genes and genomes (KEGG) database results enabled us to identify the functional categories, i.e., molecular function (16.5%), biological process (22.2%) and cellular components (61.3%). In addition, between these two species, the no hits and anonymous genes were 4.2% and 11.7% and 6.2% and 11.2% in T. laxmanni and T. angustifolia, respectively, based on the BLAST results. The study concluded that they have certain species-specific genes. Hence, the results of this study on these two species could be a valuable resource for further studies.

Molecular Cloning of Chicken Major Histocompatibility Complex Class II Molecules

  • Sung, Aree-Moon
    • Toxicological Research
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    • v.8 no.2
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    • pp.331-342
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    • 1992
  • The chicken major histocompatibility complex (MHC), the B complex, is beginning to be analyzed at the DNA level. Inbred lines of chickens have been reported to possess 3~5 MHC class II genes. To further analyzed the molecular structure of the chicken MHC class II genes, cDNA clones coding for chicken MHC class II (B-L) ${\beta}$ chain molecules were isolated from chicken spleen and liver. Tissue-specific transcription of B-L ${\beta}$genes was studied by Northern blot analysis. A high level of expression was detected for spleen poly(A)$^+$ RNA whereas a faint signal was detected for liver poly(A)$^+$ RNA. Twenty-nine cDNA clones were isolated from the spleen and eight cDNA clones were isolated from the liver. Based on restriction maps, most clones could be clustered into one family of genes. Four cDNA clones were sequenced (S7, S10 and S19 from the spleen and L1, which was identical to S19, from the liver). Complete amino acid sequences of B-L ${\beta}$ chain molecules were predicated from the nucleotide sequences of the cDNA clones. Although both the nature and the location of the conserved residues were similar in chicken and mammalian sequences, some species-specific differences were found, suggesting that the structures of the B-L molecules are similar, but not identical to their mammalian counterparts.

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Selection and identification of single-domain antibody against Peste des Petits Ruminants virus

  • Liu, Dan;Li, Lingxia;Cao, Xiaoan;Wu, Jinyan;Du, Guoyu;Shang, Youjun
    • Journal of Veterinary Science
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    • v.22 no.4
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    • pp.45.1-45.13
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    • 2021
  • Background: Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Single-domain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility. Objectives: The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV. Methods: A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2. Results: The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 106 cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein. Conclusions: The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.

Cloning of Ribulose-1,5-bisphosphate Carboxylase/oxygenase Large Subunit(rbcL) Gene from Korean Ginseng (Panax ginseng C.A. Meyer) (고려인삼(Panax ginseng C.A. Meyer) Ribulose-1,5-bisphosphate Carboxylase/oxygenase Large Subunit(rbct) Gene의 Cloning)

  • 이정헌;임용표
    • Journal of Ginseng Research
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    • v.19 no.1
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    • pp.51-55
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    • 1995
  • The DNA fragment containing ginseng ribulose-1,5-bisphosphate carboxytase/oxygenase large subunit(rbcL) gene was cloned from the ginseng chloroplast EcoRl library by colony lift hybridization with tobacco rbcL gene probe. From the screened clone, the DNA fragment containing ginseng rbcL gene was digested with several restriction enzyme and analyzed by Southern blot hybridization for the construction of restriction map. The ginseng rbcL gene fragment was subcloned in pBluescript II SK + vector and sequence analysis was performed. The nucleotide sequence of ginseng rbcL gene was compared with those of petunia, tobacco, alfalfa, rice and barley, which showed a homology of 93.1%, 95.2%, 90.5%, 85.5% and 84.3%, respectively.

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A Design of the PUMA Robot Control System Using a PC (PC를 이용한 PUMA 로봇의 제어시스템 구성)

  • Kim, Dae-Won;Lee, Won-Sik;Kyung, Kye-Hyun;Lee, Sang-Moo;Ko, Myoung-Sam;Lee, Bum-Hee
    • Proceedings of the KIEE Conference
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    • 1988.07a
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    • pp.980-985
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    • 1988
  • In this paper, a control system of the PUMA 560 robot manipulator using a PC (Personal Computer) is presented. The hardware of the designed control system is composed of IBM-PC/AT, interface board, selection board, interrupt generating circuit, and the servo control unit of the PUMA controller. A robot control library is developed using assembly and C language, and is composed of several low-level functions and arm interface routines, world model routines, arm kinematics routines, and motion command service routines. Using the designed library, joint interpolated motion and Cartesian interpolated motion of the PUMA robot manipulator are realized. In the future, our system is expected to be a very helpful basis and a useful tool for developing various control algorithms of robot manipulator using sensory information.

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Expression and Characterization of ${\alpha}$-Methylacyl CoA Racemase from Anisakis simplex Larvae

  • Kim, Bong-Jin;Kim, Sun-Mi;Cho, Min-Kyung;Yu, Hak-Sun;Lee, Yong-Seok;Cha, Hee-Jae;Ock, Mee-Sun
    • Parasites, Hosts and Diseases
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    • v.50 no.2
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    • pp.165-171
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    • 2012
  • Larval excretory-secretory products of Anisakis simplex are known to cause allergic reactions in humans. A cDNA library of A. simplex 3rd-stage larvae (L3) was immunoscreened with polyclonal rabbit serum raised against A. simplex L3 excretory-secretory products to identify an antigen that elicits the immune response. One cDNA clone, designated as ${\alpha}$-methylacyl CoA racemase (Amacr) contained a 1,412 bp cDNA transcript with a single open reading frame that encoded 418 amino acids. A. simplex Amacr showed a high degree of homology compared to Amacr orthologs from other species. Amacr mRNA was highly and constitutively expressed regardless of temperature (10-$40^{\circ}C$) and time (24-48 hr). Immunohistochemical analysis revealed that Amacr was expressed mainly in the ventriculus of A. simplex larvae. The Amacr protein produced in large quantities from the ventriculus is probably responsible for many functions in the development and growth of A. simplex larvae.

Nucleotide Sequence of Leghemoglobin cDNA from Canavalia lineata

  • An, Chung-Sun
    • Journal of Plant Biology
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    • v.37 no.2
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    • pp.167-173
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    • 1994
  • Poly(A)+ RNA was selected from Canavalia lineata root nodule RNA through oligo(dT) cellulose column and used for construction of a cDNA library using λgt10-EcoRI arms. The size of the library was 7.2$\times$105 pfu/mL. A full length leghemoglobin (Lb) cDNA clone, pCILb1(687 bp) isolated with soybean Lb probe, contained one open reading frame (ORF) of 447 bp with 54 bp plus 186 bp at 5' and 3' untranslated region, respectively. A consensus sequence of plant translation start region (AAAATGGG) was found at 5' untranslated region, and two polyadenylation-related sequence (AATAAA, AATAAG) and a conserved motif between them (gACTTGTT) were found upstream of poly(A)+ tail consisted of 13 (A)s at 3' untranslated region. The ORF encoded a polypeptide consisted of 149 amino acids with a molecular weight of 16.2 kD. Deduced amino acid sequences showed high degree of homology values with those of other Lbs ranging from 66% (Casuarina glauca) to 85% (Glycine max).

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Identification of Non-Muscle Nebulin Isoform in Human Brain Library

  • Joo, Young-Mi;Lee, Min-A;Choi, Pyung-Rak;Choi, Jae-Kyoung;Lee, Yeong-Mi;Choi, Su-Il;Kim, Myong-Shin;Jeon, Eun-Hee;Kim, So-Young;Kim, Chong-Rak
    • Biomedical Science Letters
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    • v.10 no.1
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    • pp.23-29
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    • 2004
  • Nebulin is a (Mr 600∼900 kDa) large actin-binding protein specific to skeletal muscle and thought to act as a molecular template that regulates the length of thin filaments. Cardiac muscles of higher vertebrates have been shown earlier to lack nebulin. Recently, full-length nebulin mRNA transcripts have been detected in heart muscle, but at lower levels than in skeletal muscle. Nebulin expression also was detected in the kidney, eye, and otic canal, suggesting that nebulin isoforms may also be expressed in these organs. We have searched for nebulin isoforms in brain of human using PCR and Northern blot. Here, we provide evidence that nebulin mRNA transcripts are expressed in brain. Seven nebulin isoforms (B, C, D, E, F, G and H form) are obtained in human skeletal muscle and four isoforms (B, C, G and H form) in human brain cDNA library. We cloned the 1.3 kb of nebulin fragment from human adult brain library by PCR. The identity of the PCR product was confirmed by sequence analysis. The partial brain nebulin sequence was 99% identical to the skeletal muscle cDNA as determined by Blast alignment. It contains two simple-repeats HR1, HR2 and linker-repeats exon l35∼143 except exon 140. It was different from skeletal muscle B form, which contain HR1 and HR8. These data suggest that nebulin isoform diversity occurs even more extensively than previously known, likely contributing to the distinct thin filament architecture of different striated muscles.

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