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Cloning and Expression of Lactate Dehydrogenase H Chain Gene in Adipose Tissues of Korean Cattle

  • Kim, H.H.;Seol, M.B.;Jeon, D.H.;Sun, S.S.;Kim, K.H.;Choi, Y.J.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.12
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    • pp.1670-1674
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    • 2001
  • To understand molecular mechanisms that regulate deposition and release of intramuscular fat, a fasting-induced clone was identified by differential screening from cDNA library of adipose tissues of Korean cattle. The clone had a total length of 1,319 nucleotides coding for 334 amino acids. It was identified as one encoding L-lactate dehydrogenase H chain (LDH-B). Comparison of the deduced amino acid sequences of bovine LDH-B with those of pig, human, rat, and mouse showed 98%, 98%, 97%, and 96% identity, respectively. Food deprivation for 48 h increased mRNA levels of LDH-B gene in adipose tissues of Korean cattle compared to fed- and 6 h refed- tissues. The expression of obese mRNA was examined for individual adipose tissue from several fat depots. Fasting induced expression of LDH-B gene in subcutaneous adipose tissues, but it did not affect expression levels in abdominal, perirenal and intramuscular tissues. Results demonstrate that induction of LDH-B gene during fasting may represent a metabolic shift from anaerobic state to aerobic predominance in fasted adipose tissues and that its responses to fasting are different among several adipose tissues.

Design of an Automatic Generation System of Device Drivers Using Templates (템플릿을 이용한 디바이스 드라이버 자동생성 시스템 설계)

  • Kim, Hyoun-Chul;Lee, Ser-Hoon;Hwang, Sun-Young
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.33 no.9C
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    • pp.652-660
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    • 2008
  • Applications running under embedded systems require various device drivers designed for different types and versions of the OS to manage resources effectively. In this paper, an automated device driver generator system which can generate the device drivers to be used in newer versions the target OS is proposed. In the proposed system, the structures of device drivers of specific OS are designed in the templates and stored in a library, and the target device drivers are generated by adding codes to the stored templates. Once device drivers are generated, they are registered into the kernel. The experimental results show that data transfer time has been slightly increased when compared against manually created drivers for TFT-LCD driver, USB interface keyboard/mouse driver, and AC'97 controller drivers. The code size for the generated drivers after compilation has also been increased slightly when compared with manually designed device drivers.

EST-based Survey of Gene Expression in Seven Tissue Types from the Abalone Haliotis discus hannai

  • Park, Eun-Mi;Nam, Bo-Hye;Kim, Young-Ok;Kong, Hee-Jeong;Kim, Woo-Jin;Lee, Sang-Jun;Kong, In-Soo;Choi, Tae-Jin
    • Fisheries and Aquatic Sciences
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    • v.10 no.3
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    • pp.119-126
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    • 2007
  • The analysis of expressed sequence tags (ESTs) is an efficient approach for gene discovery, expression profiling, and the development of resources for functional genomics. To analyze the transcriptome of the abalone Haliotis discus hannai, we conducted EST analysis using seven cDNA libraries made from gill, gut, hepatopancreas, skin, muscle, testis, and ovary. Redundant ESTs were assembled into overlapping contiguous sequences using the assembly program ICAtools. We found that the total 1,393 ESTs formed 135 clusters and 951 singletons, indicating that the overall redundancy of the library was 22%. Of the 1,393 clones, BLAST identified 1,278 clones (91.7%) as known genes; 115 clones (8.3%) did not match any previously described gene. Based on the major functions of their encoded proteins, the identified clones were classified into 16 broad categories. Sequence analysis revealed the presence of micro satellite-containing genes that may be valuable for further gene mapping studies. This study contributes to the identification of numerous EST clones that can be applied to further clarifying the genetics and developmental biology of abalone.

Identification of Antigenic Proteins in Trichomonas vaginalis

  • Lee, Hye-Yeon;Hyung, Su-Jin;Lee, Jong-Woong;Kim, Ju-Ri;Shin, Myeong-Heon;Ryu, Jae-Sook;Park, Soon-Jung
    • Parasites, Hosts and Diseases
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    • v.49 no.1
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    • pp.79-83
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    • 2011
  • Trichomoniasis is a sexually transmitted disease due to infection with Trichomonas vaginalis, and it can cause serious consequences for women's health. To study the virulence factors of this pathogen, T. vaginalis surface proteins were investigated using polyclonal antibodies specific to the membrane fractions of T. vaginalis. The T. vaginalis expression library was constructed by cloning the cDNA derived from mRNA of T. vaginalis into a phage ${\lambda}$ Uni-ZAP XR vector, and then used for immunoscreening with the anti-membrane proteins of T. vaginalis antibodies. The immunoreactive proteins identified included adhesion protein AP65-1, ${\alpha$-actinin, kinesin-associated protein, teneurin, and 2 independent hypothetical proteins. Immunofluorescence assays showed that AP65-1, one of the identified immunogenic clones, is prevalent in the whole body of T. vaginalis. This study led us to identify T. vaginalis proteins which may stimulate immune responses by human cells.

Decreasing effect of an anti-Nfa1 polyclonal antibody on the in vitro cytotoxicity of pathogenic Naegleria fowleri

  • Jeong, Seok-Ryoul;Kang, Su-Yeon;Lee, Sang-Chul;Song, Kyoung-Ju;Im, Kyung-Il;Shin, Ho-Joon
    • Parasites, Hosts and Diseases
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    • v.42 no.1
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    • pp.35-40
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    • 2004
  • The nfa 1 gene was cloned from a cDNA library of pathogenic Naegleria fowleri by immunoscreening; it consisted of 360 bp and produced a 13.1 kDa recombinant protein (rNfa1) that showed the pseudopodia-specific localization by immunocytochemistry in the previous study. Based on the idea that the pseudopodia-specific Nfa1 protein mentioned above seems to be involved in the pathogenicity of N. fowleri, we observed the effect of an anti-Nfa1 antibody on the proliferation of N. fowleri trophozoites and the cytotoxicity of N. fowleri trophozoites on the target cells. The proliferation of N. fowleri trophozoites was inhibited after being treated with an anti-Nfa1 polycional antibody in a dose-dependent manner for 48 hrs. By a light microscope, CHO cells co-cultured with N. fowleri trophozoites (group I) for 48 hrs showed severe morphological destruction. On the contrary, CHO cells co-cultured with N. fowleri trophozoites and anti-Nfa1 polyclonal antibody (1:100 dilution) (group II) showed less destruction. In the LDH release assay results, group I showed 50.6% cytotoxicity, and group II showed 39.3%. Consequently, addition of an anti-Nfa1 polyclonal antibody produced a decreasing effect of in vitro cytotoxicity of N. fowleri in a dose-dependent manner.

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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    • v.47 no.2
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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).

Cloning and Characterization of ${\Delta}^1$-Pyrroline-5-Carboxylate Synthetase Genes and Identification of Point Mutants in Medicago truncatula

  • Song, Ki-Hoon;Song, Dae-Hae;Lee, Jeong-Ran;Kim, Goon-Bo;Choi, Hong-Kyu;Penmetsa, R. Varma;Nam, Young-Woo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.52 no.4
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    • pp.458-468
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    • 2007
  • To tolerate environmentally adverse conditions such as cold, drought, and salinity, plants often synthesize and accumulate proline in cells as compatible osmolytes. ${\Delta}^1$-Pyrroline-5-carboxylate synthetase(P5CS) catalyzes the rate-limiting step of proline biosynthesis from glutamate. Two complete genes, MtP5CS1 and MtP5CS2, were isolated from the model legume Medicago truncatula by cDNA cloning and bacterial artificial chromosome library screening. Nucleotide sequence analysis showed that both genes consisted of 20 exons and 19 introns. Alignment of the predicted amino acid sequences revealed high similarities with P5CS proteins from other plant species. The two MtP5CS genes were expressed in response to high salt and low temperature treatments. Semi-quantitative reverse transcription-polymerase chain reaction showed that MtP5CS1 was expressed earlier than MtP5CS2, indicating differential regulation of the two genes. To evaluate the reverse genetic effects of nucleotide changes on MtP5CS function, a Targeting Induced Local Lesions in Genomes approach was taken. Three mutants each were isolated for MtP5CS1 and MtP5CS2, of which a P5CS2 nonsense mutant carrying a codon change from arginine to stop was expected to bring translation to premature termination. These provide a valuable genetic resource with which to determine the function of the P5CS genes in environmental stress responses of legume crops.

Cloning and Characterization of the pyrH Gene Encoding UMP-Kinase from Lactobacillus reuteri ATCC 55739

  • PARK JAE-YONG;NAM SU JIN;KIM JONG-HWAN;JEONG SEON-JU;KIM JUNG KON;HA YEONG LAE;KIM JEONG HWAN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.525-531
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    • 2005
  • From a genomic library of Lactobacillus reuteri ATCC 55739, one clone, NE347, carrying a pyrH gene encoding UMP kinase, was identified. pNE347 carried a 1.88 kb EcoRI fragment and the pyrH was located in the middle of the insert. pyrH ORF was 723 bp in size and capable of encoding UMP kinase composed of 240 amino acid residues. tsf encoding an elongation factor-Ts and frr encoding a ribosomal recycling factor were present upstream and downstream of pyrH, respectively. When introduced into E. coli KUR1244, a pyrH-negative strain, pNE347 restored the ability to grow at $42^{\circ}C$, indicating that pyrH from L. reuteri synthesized functional UMP kinase in E. coli. Northern blot experiment showed that pyrH and frr were cotranscribed as a 1.4 kb single transcript. pyrH was overexpressed in E. coli by using a pET26b(+) vector, and a major 25 kDa protein band appeared on SDS-polyacrylamide gel.

Criticality Analysis of KSC-4 Spent Fuel Shipping Cask (KSC-4 수송용기의 핵임계도 분석)

  • Choi, B.I.;Shin, H.S.;Park, C.M.;Ro, S.G.
    • Journal of Radiation Protection and Research
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    • v.14 no.1
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    • pp.56-65
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    • 1989
  • The nuclear criticality of the KSC-4 shipping cask which can load four assemblies of PWR spent fuel was analyzed using KENO-IV computer code and 19-group nuclear cross section set generated from 218-group neutron cross section library(DLC-43/CSRL) using AMPX system. In accordance with 10CFR71, the analysis was performed for fresh fuel assemblies, instead of the spent fuels, under both norml transportation and hypothetical accident conditions. The calculated maximum multiplication factors(Keff) of the KSC-4 cask were 0.85289 and 0.94185 for the normal transportation and hypothetical accident conditions, respectively. The highest Keff of the KSC-4 cask is within the subcritical limit prescribed in l0CFR71 and accordingly the KSC-4 cask is safely designed in terms of nulear criticality.

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VLSI architecture design of CAVLC entropy encoder/decoder for H.264/AVC (H.264/AVC를 위한 CAVLC 엔트로피 부/복호화기의 VLSI 설계)

  • Lee Dae-joon;Jeong Yong-jin
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.30 no.5C
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    • pp.371-381
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    • 2005
  • In this paper, we propose an advanced hardware architecture for the CAVLC entropy encoder/decoder engine for real time video compression. The CAVLC (Context-based Adaptive Variable Length Coding) is a lossless compression method in H.264/AVC and it has high compression efficiency but has computational complexity. The reference memory size is optimized using partitioned storing method and memory reuse method which are based on partiality of memory referencing. We choose the hardware architecture which has the most suitable one in several encoder/decoder architectures for the mobile devices and improve its performance using parallel processing. The proposed architecture has been verified by ARM-interfaced emulation board using Altera Excalibur and also synthesized on Samsung 0.18 um CMOS technology. The synthesis result shows that the encoder can process about 300 CIF frames/s at 150MHz and the decoder can process about 250 CIF frames/s at 140Mhz. The hardware architectures are being used as core modules when implementing a complete H.264/AVC video encoder/decoder chip for real-time multimedia application.