• Title/Summary/Keyword: DNA coding.

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Simultaneous Expression of Pseudomonas sp. Endo-1,4$\beta$-Glucanase and $\beta$-1,4=Glucisidase Gene in Escherichia coli and Saccharomyces cerevisiae (Pseudomonas sp. Endo-1,4-$\beta$-Glucanase와 $\beta$-1,4-Glucosidase 유전자의 대장균 및 효모에서의 동시 발현)

  • Kim, Yang-Woo;Chun, Sung-Sik;Chung, Young-Chul;Sung, Nack-Kie
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.652-658
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    • 1995
  • We attempted simultaneous expression of genes coding for endoglucanase and $\beta $-glucosidase from Pseudomonas sp. by using a synthetic two-cistron svstem in Escherichia coli and Saccharomyces cerevisiae. Two-cistron system, 5'--tac promoter-endoglucanase gene--$\beta $-glucosidase gene-- 3', 5'-tac promoter--$\beta $-glucosidase gene--endoglucanase gene--3' and 5'-tac promoter--endoglucanase gene--SD sequence--$\beta $-glucosidase gene--3, were constructed, and expressed in E. coli and S. cerevisiae. The E. coli and S. cerevisiae contained two-cistron system produced simultaneously endoglucanase and $\beta $-glucosidase. The recombinant genes contained the bacterial signal peptide sequence produced low level of endoglucanase and $\beta $-glucosidase in S. cerevisiae transformants: Approximately above 44% of two enzymes was localized in the intracellular fraction. The production of endoglucanase and $\beta $-glucosidase in veast was not repressed in the presence of glucose or cellobiose. The veast strain contained recombinant DNA with two genes hydrolyzed carboxvmethyl cellulose, and these endoglucanase and $\beta $-glucosidase degraded CMC synergistically to glucose, cellobiose and oligosaccharide. This result suggests the possibility of the direct bioconversion of cellulose to ethanol by the recombinant yeast.

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Molecular Cloning and the Nucleotide Sequence of a Bacillus sp. KK-l $\beta$-Xylosidase Gene

  • Chun, Yong-Chin;Jung, Kyung-Hwa;Lee, Jae-Chan;Park, Seung-Hwan;Chung, Ho-Kwon;Yoon, Ki-Hong
    • Journal of Microbiology and Biotechnology
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    • v.8 no.1
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    • pp.28-33
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    • 1998
  • A gene coding for ${\beta}$-xylosidase from thermophilic xylanolytic Bacillus sp. KK-1 was cloned into Escherichia coli using plasmid pBR322. Recombinant plasmid DNAs were isloated from E. coli clones which were capable of hydrolyzing 4-methylumbelliferyl-${\beta}$-D xylopyranoside. Restriction analysis showed the DNAs to share a common insert DNA. Xylo-oligosaccharides, including xylotriose, xylotetraose, xylopentaose, and xylobiose were hydrolyzed to form xylose as an end product by cell-free extracts of the E. coli clones, confirming that the cloned gene from strain KK-1 is ${\beta}$-xylosidase gene. The ${\beta}$-xylosidase gene of strain KK-1 designated as xylB was completely sequenced. The xylB gene consisted of an open reading frame of 1,602 nucleotides encoding a polypeptide of 533 amino acid residues, and a TGA stop codon. The 3' flanking region contained one stem-loop structure which may be involved in transcriptional termination. The deduced amino acid sequence of the KK-1 ${\beta}$-xylosidase was highly homologous to the ${\beta}$-xylosidases of Bacillus subtilis and Bacillus pumilus, but it showed no similarity to a thermostable ${\beta}$-xylosidase from Bacillus stearothermophilus.

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Homology Analysis Among the Biphenyl and 4-Chlorobiphenyl Degrading Genes by Southern Hybridization (Southern Hybridization에 의한 Biphenyl 및 4-Chlorobiphenyl 분해유전자들의 상동성 분석)

  • 남정현;김치경;이재구;이길재
    • Microbiology and Biotechnology Letters
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    • v.22 no.1
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    • pp.37-44
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    • 1994
  • The homology among the genes coding for degradation of bipheny(BP) and 4-chlorobiphenyl(4CB) was comparatively analyzed by Southern hybridization in several BP/4CB degrading bacterial strains. As the hybridization results of their genomic DNAs with pcbABCD as the DNA probe, the group of Pseudomonas sp. DJ-12. P08 and P27 strain was separated by the group of P20 and P1242 strains. The P. pseudoalcaligenes KF707 showed the hybidization signal which was homologous to the group of DJ-12, but they had different restriction endonuclease sites. The pcbAB genes in pCUl recombinant plasmid from Pseudomonas sp. DJ-12 appeared to be homologous to pchAB genes in pKTF20 cloned from P. pseudoalcaligenes KF707, but the C genes in both strains were not homologous. The bphABC in pKTF20 showed the signals homologous to the cbp ACB in pAW6194 cloned from P. putida OU83, but homologous signal was not found botween the pcbABCD genes in pCUl and the cbpADCB genes in pAW6194 recombbinant plasmid.

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Plastid genome of Aster altaicus var. uchiyamae Kitam., an endanger species of Korean asterids

  • Park, Jihye;Shim, Jaekyung;Won, Hyosig;Lee, Jungho
    • Journal of Species Research
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    • v.6 no.1
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    • pp.76-90
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    • 2017
  • Aster altaicus var. uchiyamae Kitam. is an endemic taxon of Korea and is protected by law as an endanger taxon. The genetic information of A. altaicus var. uchiyamae is unavailable in Genbank. Here we sequenced chloroplast genome of A. altaicus var. uchiyamae. The cp-genome of Aster altaicus var. uchiyamae was 152,446 bps in size: LSC was 84,240 bps, IR 25,005 bps, SSC 18,196 bps. The cp-genome contains 112 genes and 21 introns consisted of 79 protein coding genes(PCGs), 4 RNA genes, and 29 tRNA genes, with 20 group II introns and one group I intron. There were three pseudo-genes including ${\psi}$-ycf1, ${\psi}$-rps19, and ${\psi}$-trnT_GGU. Eighteen genes, five introns, and parts of two genes and an intron are found within the IR, which has two copies. The cp-DNA of Aster altaicus var. uchiyamae is distinguished from A. spathulifolius, only known cp-genome of the genus Aster, by 172 SNP in genic regions of 43 PCGs and 21 indels in 11 PCGs and SSU. The chloroplast genome sequence was deposited at GenBank (KX35265).

Characterization of Spodoptera exigua Nuclear Polyhedrosis Virus Polyhedrin Gene Structure (파밤나방 핵다각체병 바이러스의 다각체 단백질 유전자 구조)

  • 최재영;김우진
    • Journal of Sericultural and Entomological Science
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    • v.38 no.2
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    • pp.144-149
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    • 1996
  • To develope the baculovirus expression vector system (BEVS) using Spodoptera exigua nuclear polyhedrosis virus (SeNPV), we characterized the polyhedrin of SeNPV. The SeNPV polyhedra was irregular and composed of the major protein molecular weight of 30 kDa determined by electronmicroscopy and SDS-AGE analysis, respectively. The nucleotid suquences of 876 bases including the coding region of polyhedrin gene was determined and it was revealed that the polyhedrin gene is located within Xho I 3.0Kb and Nco I 6.0 Kb by Southern blot analysis, respectively. Also, the Xho I 3.0 Kb and the Nco I 6.0 Kb fragments were cloned and restriction enzyme map of these clones were determined.

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Phenylalanine Ammonia-Lyase Gene (NtPAL4) Induced by Abiotic Stresses in Tobacco (Nicotiana tabacum)

  • Han, Woong;Wang, Myeong-Hyeon
    • Korean Journal of Plant Resources
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    • v.23 no.6
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    • pp.535-540
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    • 2010
  • Phenylalanine ammonia-lyase (PAL), a key enzyme of the phenylpropanoid biosynthesis pathway, is activated by a number of developmental and environmental cues. The coding region of the NtPAL4 gene was 2,154 bp in length, and its deduced protein was composed of 717 amino acids. Sequence analysis of NtPAL4 cDNA from tobacco (Nicotiana tabacum L.) revealed high structural similarity to PAL genes of other plant species. The NtPAL4 gene exists as a single copy in the tobacco plant, and its transcripts were strongly expressed in flowers and leaves. NtPAL4 expression was significantly induced in response to NaCl, mannitol, and cold treatments, but it was not induced by abscisic acid (ABA). NtPAL4 expression decreased gradually after treatment with ABA and $H_2O_2$; however, NtPAL4 transcripts accumulated after treatment with methyl viologen (MV). Our results suggest that the NtPAL4 gene may function in response to abiotic stresses.

Genetic Risk Prediction for Normal-Karyotype Acute Myeloid Leukemia Using Whole-Exome Sequencing

  • Heo, Seong Gu;Hong, Eun Pyo;Park, Ji Wan
    • Genomics & Informatics
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    • v.11 no.1
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    • pp.46-51
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    • 2013
  • Normal-karyotype acute myeloid leukemia (NK-AML) is a highly malignant and cytogenetically heterogeneous hematologic cancer. We searched for somatic mutations from 10 pairs of tumor and normal cells by using a highly efficient and reliable analysis workflow for whole-exome sequencing data and performed association tests between the NK-AML and somatic mutations. We identified 21 nonsynonymous single nucleotide variants (SNVs) located in a coding region of 18 genes. Among them, the SNVs of three leukemia-related genes (MUC4, CNTNAP2, and GNAS) reported in previous studies were replicated in this study. We conducted stepwise genetic risk score (GRS) models composed of the NK-AML susceptible variants and evaluated the prediction accuracy of each GRS model by computing the area under the receiver operating characteristic curve (AUC). The GRS model that was composed of five SNVs (rs75156964, rs56213454, rs6604516, rs10888338, and rs2443878) showed 100% prediction accuracy, and the combined effect of the three reported genes was validated in the current study (AUC, 0.98; 95% confidence interval, 0.92 to 1.00). Further study with large sample sizes is warranted to validate the combined effect of these somatic point mutations, and the discovery of novel markers may provide an opportunity to develop novel diagnostic and therapeutic targets for NK-AML.

Functional Analysis of Pepper Cys2/His-Type Zinc-Finger Protein Promoter Region in Response to Bacterial Infection and Abiotic Stresses in Tobacco Using Agrobacterium-Mediated Transient Assay

  • Kim, Sang-Hee;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.21 no.1
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    • pp.39-46
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    • 2005
  • The promoter region flanking the 5’ CAZFP1 coding region was isolated from the genomic DNA of Capsicum annuum. To identify the upstream region of the CAZFP1 gene required for promoter activity, a series of CAZFP1 promoter deletion derivatives was created. Each deletion construct was analyzed by Agrobacterium-mediated transient transformation in tobacco leaves after infection by Pseudomonas syringae pv. tabaci, or treatment with methyl jasmonate (MeJA), ethylene, abscisic acid (ABA), salicylic acid (SA), cold and wounding. Promoter fragments of 685 bp or longer showed 7-fold or greater induction after P. s. pv. tabaci infection and MeJA treatment. The CAZFP1 full-length promoter (-999 bp) also showed 6-fold induction in response to ethylene. The transiently transformed tobacco leaves with the CAZFP1 full length promoter fused-GUS gene showed more than 5-fold induction in response to SA, ABA and cold. These results suggest that the CAZFP1 promoter contains responsive elements for pathogen, MeJA, ethylene, SA, ABA and cold.

Genomic Structure of the Luciferase Gene of the Firefly, Lampyris noctiluca

  • Jianhong Li;Park, Yong-Soo;Lee, Sang-Chul;Lee, Sang-Mong;Kim, Jong-Gill;Kim, Iksoo;Kim, Keun-Young;Sohn, Hung-Dae;Jin, Byung-Rae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.7 no.2
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    • pp.155-159
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    • 2003
  • We describe here the complete nucleotide sequence and the exon-intron structure of the luciferase gene of the firefly, Lampyris noctiluca. The luciferase gene of the L. noctiluca firefly consisted of six introns and seven exons coding for 547 amino acid residues. From the translational start site to the end of last exon, the genomic DNA length of the L. noctiluca luciferase gene spans 1,976 bp.

Complete Nucleotide Sequence of Cytochrome P450 2El Expressed in the Rat Brain

  • Shin Song Woo;You Kwan Hee;Ryu Hye Myung;Kim Su Won;Kwon Oh Sik;Song Jae Chan;Kim Myoung Hee;Kim Dae Ran;Yoo Min
    • Biomedical Science Letters
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    • v.11 no.2
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    • pp.109-113
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    • 2005
  • From the RT-PCR amplifications using mRNA templates isolated from Sprague-Dawley rat brains, we isolated a cDNA fragment of 1,524 bp which covered the full coding information of the rat brain CYP2El. Its nucleotide sequence was identical to the previously reported rat liver CYP2El mRNA except for the difference of one base (A to C at the nucleotide position 73). This difference also altered the amino acid Lys to GIn. However, no insertion or deletion of nucleotide(s) which could alter the reading frame was found within the structure of this rat brain CYP2El. This study should provide the molecular basis regarding the pathophysiological function of CYP2El in the brain.

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