• Title/Summary/Keyword: Delta encoding

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Proline accumulation and transcriptional regulation of proline biothesynthesis and degradation in Brassica napus

  • Xue, Xingning;Liu, Aihua;Hua, Xuejun
    • BMB Reports
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    • v.42 no.1
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    • pp.28-34
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    • 2009
  • To understand the molecular mechanism underlying proline accumulation in Brassica napus, cDNAs encoding ${\Delta}^1$-pyrroline-5-carboxylate synthetase (BnP5CS), ornithine $\delta$-aminotransferase (BnOAT) and proline dehydrogenase (BnPDH) were isolated and characterized. Southern blot analysis of BnP5CSs in B. napus and its diploid ancestors suggested a gene loss may have occurred during evolution. The expression of BnP5CS1 and BnP5CS2 was induced, while the expression of BnPDH was inhibited under salt stress, ABA treatment and dehydration, prior to proline accumulation. The upregulation of BnOAT expression was only detected during prolonged severe osmotic stress. Our results indicate that stress-induced proline accumulation in B. napus results from the reciprocal action of activated biosynthesis and inhibited proline degradation. Whether the ornithine pathway is activated depends on the severity of stress. During development, proline content was high in reproductive organs and was accompanied by markedly high expression of BnP5CS and BnPDH, suggesting possible roles of proline during flower development.

Invesigation of Functional Roles of a Protein Kinase in a Fungal Plant Pathogen, Magnaporthe oryzae

  • Han, Joon-Hee;Shin, Jong-Hwan;Kim, Kyoung Su
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.43-43
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    • 2014
  • The rice blast disease caused by of Magnaporthe oryzae is one of the most destructive diseases of rice. By the microarray analysis, we profiled expression changes of genes during conidiation and found out many putative genes that are up-regulated. Among those, we first selected MGG_06399 encoding a dual-specificity tyrosine-regulated protein kinase (DYRK), homologous to YAK1 in yeast. To investigate functional roles of MoYAK1, We made ${\Delta}Moyak1$ mutants by homology dependent gene replacement. The deletion mutant showed a remarkable reduction in conidiation and produced abnormally shaped conidia smaller than those of wild type. The conidia form ${\Delta}Moyak1$ were able to develop a germ tube, but failed to form apppressoria on a hydrophobic coverslip. The ${\Delta}Moyak1$ formed appressria on a hydrophobic cover slip when exogenous cAMP was induced, but the appressoria shape was abnormal. The ${\Delta}Moyak1$ also formed appressoria abberent in shape on onion epidermis and rice sheaths and failed to penetrate the surface of the plants. These data indicate that MoYAK1 is associated with cAMP/PKA pathway and important for conidiation, appressorial formation and pathogenic development in Magnaporthe oryzae. Detailed characterization of MoYAK1 will be presented.

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Endoproteolytic Processing of Human Parathyroid Hormone in Saccharomyces cerevisiae Mutants Lacking Genes Encoding the Yap3, Mkc7, and Kex2 Endoproteases

  • Choi, Won-A.;Kang, Hyun-Ah;Chung, Bong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.679-682
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    • 1999
  • When the human parathyroid hormone (hPTH) is expressed as a secretory product in S. cerevisiae, most of the secreted hPTH is internally cleaved by endoproteolytic processing. To investigate whether the yeast endoproteases such as Kex2p, Yap3p, and Mkc7p are involved in the endoproteolytic processing of hPTH in S. cerevisiae, hPTH was expressed in S. cerevisiae mutants deficient in one or two of the following well-known endoproteases such as Kex2p, Mkc7p, and Yap3p. Among these mutants, the yap3-disrupted(yap3$\Delta$) and yap3/mkc7-disrupted (yap3Δmkc7$\Delta$) yeasts showed a significant reduction in the extent of hPTH proteolysis. In contrast, the mkc7-disrupted (mkc7$\Delta$) yeast did not reduce the proteolysis of hPTH as compared to the wild type. This suggests that Mkc7p is not involved in the endoproteolytic processing of hPTH. It was also found that the kex2-disrupted (kex2$\Delta$) mutant was not able to secrete a detectable amount of hPTH.

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Purification and Characterization of the N-terminally Truncated DNA Polymerase from Thermus thermophilus HJ6 (Thermus thermophilus HJ6 유래 N-말단 결실 DNA Polymerase의 정제 및 특성)

  • Jeon, Sung-Jong;Seo, Min-Ho
    • Microbiology and Biotechnology Letters
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    • v.38 no.2
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    • pp.158-162
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    • 2010
  • The gene encoding N-terminally truncated Tod polymerase ($\Delta$Tod polymerase) from Thermus thermophilus HJ6 was expressed in Escherichia coli under the control of the lambda pR and pL tandem promoters on the expression vector pJLA503. The N-terminal domain (250 amino acids) of Tod polymerase was removed without significant effect on enzyme activity and stability, while no 5'$\rightarrow$3' exonuclease activity was detected. The $\Delta$Tod polymerase was verified to possess very efficient reverse transcriptase (RT) activity in the presence of $MgCl_2$. The cDNA can also be amplified in the polymerase chain reaction (PCR) with this mutant enzyme. The $\Delta$Tod polymerase was exhibited higher activity than the Taq polymerase in a one-step RT-PCR.

Cloning and Strong Expression of a Bacillus subtilis WL-3 Mannanase Gene in B. subtilis

  • Yoon, Ki-Hong;Lim, Byung-Lak
    • Journal of Microbiology and Biotechnology
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    • v.17 no.10
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    • pp.1688-1694
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    • 2007
  • A gene encoding the mannanase of Bacillus subtilis WL-3, which had been isolated from Korean soybean paste, was cloned into Escherichia coli and the nucleotide sequence of a 2.7-kb DNA fragment containing the mannanase gene was subsequently determined. The mannanase gene, designated manA, consisted of 1,080 nucleotides encoding a polypeptide of 360 amino acid residues. The deduced amino acid sequence was highly homologous to those of mannanases belonging to glycosyl hydrolase family 26. The manA gene was strongly expressed in B. subtilis 168 by cloning the gene downstream of a strong B. subtilis promoter of plasmid $pJ27{\Delta}88U$. In flask cultures, the production of mannanase by recombinant B. subtilis 168 reached maximum levels of 300 units/ml and 450 units/ml in LB medium and LB medium containing 0.3% locust bean gum, respectively. Based on the zymogram ofthe mannanase, it was found that the mannanase produced by recombinant B. subtilis could be maintained stably without proteolytic degradation during the culture time.

Regulation of fpr Gene Encoding NADPH : Ferredoxin Oxidoreductase by the soxRS Locus in Escherichia coli

  • Koh, Young-Sang;Choih, Jenny;Roe, Jung-Hye
    • Journal of Microbiology
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    • v.34 no.2
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    • pp.137-143
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    • 1996
  • We isolated a promoter inducible by paraquat, a superoxide-generating agent, from Escherichia coli using a promoter-probing plasmid pRS415. From sequence analysis we found out the promoter is for fpr ENCODING nadph : ferredoxin oxidoreductase. We constructed on operon fusion of lacZ gene with fpr promoter to monitor the expression of the gene in the single-copy state. LacZ expression generators, menadione and plumbagin, also induced the expression of .betha.-galactosidase in the fusion strain. On the other hand, no significant induction was observed by treatment with hydrogen peroxide, ethanol, and heat shock. Induction of .betha.-galactosidase was significantly reduced by introducing a .DELTA. sox 8 :: cat of soxS3 :: Tn10 mutation into the fusion strain, indicating that fpr gene is a member of the soxRS regulon. The transcriptional start site was determined by primer extension analysis. Possible roles of fpr induction in superoxide stress were discussed.

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Expression and Purification of Delta Sleep-Inducing Peptide in Escherichia coli

  • Oh, Kwang-Seok;Na, Do-Kyung;Kweon, Mee-Hyang;Sung, Ha-Chin
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.620-623
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    • 2003
  • The delta sleep-inducing peptides (DSIP, Trp-Ala-Gly-Gly-Asp-Ala-Ser-Gly-Glu) is an important regulatory hormone, controlling hypothalamus and pituitary functions. In the current study, an expression system was designed for the rapid and economic expression oi recombinant DSIP for biophysical studies. Artificially synthesized oligonucleotides encoding DSIP were cloned into a pGEX-KG vector and expressed in E. coli (BL21). The recombinant GST-DSIP was then readily purified using a GST affinity column. To obtain intact DSIP from the GST-DSIP, thrombin cleavage and a CNBr reaction were successively carried out. The DSIP in the CNBr reaction mixture was subjected to RP-HPLC purification to yield 1.2 mg DSIP from a 1 liter culture of E. coli. Identification of the DSIP was peformed using MALDI-MS and an amino acid composition analysis.

Simultaneous and Sequential Integration by Cre/loxP Site-Specific Recombination in Saccharomyces cerevisiae

  • Choi, Ho-Jung;Kim, Yeon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.826-830
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    • 2018
  • A Cre/loxP-${\delta}$-integration system was developed to allow sequential and simultaneous integration of a multiple gene expression cassette in Saccharomyces cerevisiae. To allow repeated integrations, the reusable Candida glabrata MARKER (CgMARKER) carrying loxP sequences was used, and the integrated CgMARKER was efficiently removed by inducing Cre recombinase. The XYLP and XYLB genes encoding endoxylanase and ${\beta}$-xylosidase, respectively, were used as model genes for xylan metabolism in this system, and the copy number of these genes was increased to 15.8 and 16.9 copies/cell, respectively, by repeated integration. This integration system is a promising approach for the easy construction of yeast strains with enhanced metabolic pathways through multicopy gene expression.

A Study on the Design of Multiplierless FIR Filters (승산기를 사용하지 않는 FIR필터의 설계에 관한 연구)

  • 신재호;이종옥
    • Journal of the Korean Institute of Telematics and Electronics
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    • v.22 no.3
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    • pp.16-22
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    • 1985
  • In this paper a new algorithm named modified delta modulation (MDM) for encoding filter coefficients is proposed. And this paper presents the designing method of multiplier less FIR filters rosin영 Proposed MDM a19orithm. In the delta modulation (DM) system the quantiaation levels consist of two levels $\pm$1, but in newly proposed MDM algorithm quantization levels are extended to many levels 0, $\pm$2$^n$, n=0, 1, 2... It is recognized by the result of computer simulations that frequency response of multi-plierless FIR filters designed by MDM algorithm is relatively good. And comparing with con-ventional FIR filters on the number of hardware devices, this filter needs a little increased memory, but regardless of filter order it needs only one multiplier which is used for signal scaling.

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A Fast Rough Mode Decision Algorithm for HEVC

  • Yao, Wei-Xin;Yang, Dan;Lu, Gui-Fu;Wang, Jun
    • Journal of Information Processing Systems
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    • v.15 no.3
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    • pp.492-499
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    • 2019
  • HEVC is the high efficiency video coding standard, which provides better coding efficiency contrasted with the other video coding standard. But at the same time the computational complexity increases drastically. Thirty-five kinds of intra-prediction modes are defined in HEVC, while 9 kinds of intra prediction modes are defined in H.264/AVC. This paper proposes a fast rough mode decision (RMD) algorithm which adopts the smoothness of the up-reference pixels and the left-reference pixels to decrease the computational complexity. The three step search method is implemented in RMD process. The experimental results compared with HM13.0 indicate that the proposed algorithm can save 39.7% of the encoding time, while Bjontegaard delta bitrate (BDBR) is increased slightly by 1.35% and Bjontegaard delta peak signal-to-noise ratio (BDPSNR) loss is negligible.