• Title/Summary/Keyword: DNA coding.

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Cloning and Gene Manipulation of a cDNA Encoding A Cellulase Produced by the Mulberry Longicorn Beetle, Apriona germari

  • Kim, Yong-Soon;Lee, Sung-Jin;Sohn, Bong-Hee;Kang, Pil-Don;Lee, Sang-Uk;Jin, Byung-Rae
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.10a
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    • pp.157-159
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    • 2003
  • We have cloned and sequenced the cDNA coding fur a cellulase from the mulberry longicorn beetle, Apriona germari, with the polymerase chain reaction. And then we have constructed the recombinant plasmid vector for Bombyx mori transfomation experiment. (omitted)

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Purification, Characterization, and Cloning of Trimethylamine Dehydrogenase from Methylophaga sp. Strain SK1

  • Kim, Hee-Gon;Kim, Yan;Lim, Heon-Man;Shin, Hyun-Jae;Kim, Si-Wouk
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.4
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    • pp.337-343
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    • 2006
  • Trimethylamine dehydrogenase (TMADH, EC 1.5.99.7), an iron-sulfur flavoprotein that catalyzes the oxidative demethylation of trimethylamine to form dimethylamine and formaldehyde, was purified from Methylophaga sp. strain SK1. The active TMADH was purified 12.3-fold through three purification steps. The optimal pH and temperature for enzyme activity was determined to be 8.5 and $55^{\circ}C$, respectively. The $V_{max}\;and\;K_m$ values were 7.9 nmol/min/mg protein and 1.5 mM. A genomic DNA of 2,983 bp from Methylophaga sp. strain SK1 was cloned, and DNA sequencing revealed the open reading frame (ORF) of the gene coding for TMADH. The ORF contained 728 amino acids with extensive identity (82%) to that of Methylophilus methylotrophus $W_3A_1$.

Enhanced stability of Pseudomonas sp. Endo-1,4-$\beta$$\beta$-1,4-Glucosidase Gene (Pseudomonas sp. 유래 Endo-1,4-$\beta$-Glucanase 및$\beta$-1,4-Glucosidase 유전자의 안정성 개선)

  • Kim, Yang-Woo;Chun, Sung-Sik;Chung, Young-Chul;Roh, Jong-Soo;Sung, Nack-Kie
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.659-664
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    • 1995
  • To improve stability of recombinant DNA pLC1 encoding endoglucanase gene and pGL1 encoding $\beta $-glucosidase gene, DNA fragments of genes coding endoglucanase and $\beta $-glucosidase were cloned within the recA gene on a pDR1453, and the pDRE10 and pDRG20 of recombinant plasmids were integrated into the recA gene on the E. coli 1100 chromosomal DNAs. The stability of inheritance was completely maintained in E. coli 1100; Transformants E. coli 1100/pDREIO and pDRG20 were expressed well by recA promoter and increased endoglucanase and $\beta $-glucosidase activities. This method can be used as a model to improve the stability of recombinant plasmid in large scale culture.

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Molecular Cloning of $\beta$-Galactosidase from Bacillus subtilis HP-4

  • Kim, Jeong-Ho;Lee, Jae-Chang;Huh, Jeong-Won;Chung, Ki-Chul
    • Journal of Microbiology and Biotechnology
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    • v.1 no.4
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    • pp.227-231
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    • 1991
  • A gene coding for a $\beta$-galactosidase of Bacillus subtilis HP-4 was cloned in E. coli JM109 by inserting HindIII digested fragment of B. subtilis HP-4 chromosomal DNA into the site of pBR322 and selecting recombinant transformant showing blue color on X-gal plate. The recombinant plasmid, named pBG109, was found to contain the 1.4 Kbp HindIII fragment originated from B. subtilis HP-4 chromosomal DNA by Southern hybridization. The cloned gene was stably maintained and expressed in E. coli JM109 and the pBG109 encoded $\beta$-galactosidase had the same enzymatic properties as those of $\beta$-galactosidase produced by B. subtilis HP-4.

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Cloning and expression of glutathione S-transferase (GST) cDNA from Gossypium hirsutum L.

  • Kang, Won-Hee;Kim, Myong-Jo;Lim, Jung-Dae;Yun, Song-Joong;Chung, Ill-Min;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.4
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    • pp.294-297
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    • 2002
  • A gene coding for the GST of cotton (Gh-5) was cloned into Escherichia coli and experssed. The enzyme remained within the cytoplasm of E. coli. An 696 bp open reading frame was in the 988 base pair fragment of the recombinant plasmid pET-30b(+). The deduced protein sequence consists of 232 amino acids and has a molecular mass of 30235.58 Da. The cloned enzyme conjugated reduced glutathione and 1-chloro-2,4-dinitrobenzene (CDNB). Plant GST cDNA was expressed in microbe and produced polypeptide had function as an enzyme.

A cDNA Clone for the 5' Exon of Chloroplast ATP Synthase Subunit I Gene (atpF) from Broccoli (Brassica oleracea L. var. Italica) and Its Expression Pattern

  • Choo Bong Hong
    • Journal of Plant Biology
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    • v.38 no.2
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    • pp.137-141
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    • 1995
  • We isolated a cDNA clone, BLSC1, encoding 5' exon of ATP synthase CF0 subunit I from broccoli. BLSC1 is 285 nucleotides long which consists of a 5' noncoding region of 34 nucleotides, a 5' exon of 145 nucleotides and an intron of 106 nucleotides. The 5' exon codes for 48 amino acids which reveals mostly hydrophobic. The amino acid sequence deduced from BLSC1 shares 83%, 83% and 91% identities with the genes coding for atpF from wheat, rice and spinach, respectively. Genomic Southern blot analysis for BLSC1 showed a typically strong signal for a gene located in the chloroplast genome. Northern blot analysis identified three major classes of transcripts showing strong positive signals in the leaves, but only trace amounts of the transcripts were identified in the other organs like stems, flowr buds and roots.

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Influence of Toxicologically Relevant Metals on Human Epigenetic Regulation

  • Ryu, Hyun-Wook;Lee, Dong Hoon;Won, Hye-Rim;Kim, Kyeong Hwan;Seong, Yun Jeong;Kwon, So Hee
    • Toxicological Research
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    • v.31 no.1
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    • pp.1-9
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    • 2015
  • Environmental toxicants such as toxic metals can alter epigenetic regulatory features such as DNA methylation, histone modification, and non-coding RNA expression. Heavy metals influence gene expression by epigenetic mechanisms and by directly binding to various metal response elements in the target gene promoters. Given the role of epigenetic alterations in regulating genes, there is potential for the integration of toxic metal-induced epigenetic alterations as informative factors in the risk assessment process. Here, we focus on recent advances in understanding epigenetic changes, gene expression, and biological effects induced by toxic metals.

Cloning and Expression of a cDNA AAPT3 Encoding Aminoalcoholphosphotransferase Isoform from Chinese Cabbage

  • Kim, Kwang-Soo;Park, Jong-Ho;Cho, Sung-Ho
    • Animal cells and systems
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    • v.8 no.2
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    • pp.105-109
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    • 2004
  • Aminoalcoholphosphotransferase catalyzes the synthesis of phosphatidylcholine and phosphatidylethanolamine from diacylglycerol plus a CDP-aminoalcohol such as CDP-choline or CDP-ethanolamine. Previously we suggested the presence of possible isoforms of this enzyme from Chinese cabbage roots and now report the cDNA cloning and expression analysis of AAPT3 encoding a third isoform of aminoalcoholphosphotransferase (AAPT3). AAPT3 contains an open reading frame of 1,176 bp coding for a protein of 392 amino acids. It shares 96 and 95% identity with Chinese cabbage AAPT1 and AAPT2, respectively, at the deduced amino acid level. The results from reverse transcriptase-polymerase chain reaction analysis indicate that expression of AAPT3 is up-regulated by low temperature as well as AAPT1 and AAPT2.

Molecular Cloning and Characterization of Neuronal $\beta$-subunit of Large-Conductance$Ca^{2+}$-activated $K^+$ Channels from Rat Brain

  • Heo, Moon-Sun;Ha, Tal-Soo;Park, Chul-Seung
    • Proceedings of the Korean Biophysical Society Conference
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    • 2001.06a
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    • pp.38-38
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    • 2001
  • We cloned the cDNA encoding the neuron-specific $\beta$-subunit ($\beta$4) of large-conductance calcium-activated potassium channels from rat brain and determined the DNA sequences of the entire coding region (GenBank accession; AY028605). The deduced amino acid sequences of r$\beta$4, 210 amino acids in length, are closely related to the $BK_{Ca}$ $\beta$4 subunits of other species but show only limited sequence homology to other $\beta$-subunits, $\beta$1-$\beta$3.(omitted)d)

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Cloning of the gense coding for extracellular proteases from alkalophilic xanthomonas SP. JK311

  • Kim, Young-Hun;Jang, Ji-Yeon;Yeehn Yeeh;Kim, Yong-Ho;Kim, Sang-Hae
    • Journal of Microbiology
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    • v.33 no.4
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    • pp.344-349
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    • 1995
  • The alkalophilic bacterium, Xanthomonas sp. JK311, producing extracellular proteases, was isolated from soil. Xanthomonas sp. JK311 produced five extracellular proteases that are all metalloproteases. Four of them were resistant against 1% SDS. Chromosomal DNA of the Xanthomonas sp. JK311 was digested with BamHI and cloned into PUC19. Among E. coli strain HB101 transformants, a clone secreting the proteases was screened through halo formation on skim-milk agar plate and by Southern blot analysis. It had the recombinant plasmid pXEP-1 containing the 7.5 kb-BamHI DNA fragment and produced three extacellular proteases. Their protease properties corresponded to those of Xanthomonas sp. JK311.

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