• Title/Summary/Keyword: ITS gene sequence

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Location and Nucleotide Sequence of the Bombyx mori Nuclear Polyhedrosis Virus Polyhedrin Gene (누에 핵다각체병 바이러스의 다각체 단백질 유전자의 위치 탐색 및 염기서열)

  • 우수동;김현욱;박범석;강석권;양재명;정인식
    • Journal of Sericultural and Entomological Science
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    • v.34 no.2
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    • pp.20-25
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    • 1992
  • The location of the polyhedrin gene of Bmbyx mori nuclear polyhedrosis virus(BmNPV) was determined by using a cloned polyhedrin gene from the Autographa californica nuclear polyhedrosis virus(AcNPV) as a hybridization probe. The 7.4 Kb PstⅠ fragment DNA of Bm-NPV was cloned to plasmid pUC19 vector. A fragment containing this gene was mapped and sequenced in its entire polyhedrin reading frame. Nucleotide sequences comparison of the polyhedrin of the BmNPV to that of previously reported by Ⅰatrou(1985) revealed that the sequence varied in 10 base, Comparison of the amino acid sequence of the two structured gene revealed that coding sequence varied 74 valine to isoleucine, 76 aspargine to serine and 155 methionine to valine.

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Molecular Characterization of a Chinese cabbage cDNA, C-DH, Predominantly Induced by Water-Deficit Stress and Plant Hormone, ABA (수분부족 및 식물호르몬, ABA에 의하여 발현이 유도되는 배추의 C-DH cDNA에 대한 분자적 특성)

  • 정나은;이균오;홍창휘;정배교;박정동;이상열
    • Korean Journal Plant Pathology
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    • v.14 no.3
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    • pp.240-246
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    • 1998
  • A cDNA encoding desiccation-related protein was isolated from a flower bud cDNA library of Chinese cabbage (C-DH) and its nucleotide sequence was characterized. It contains 679 bp nucleotides with 501 bp open reading frame. The amino acid sequence of the putative protein showed the highest amino acid sequence homology (79 % identity) to dehydrin protein in Gossypium hirsutum. Also, the C-DH shares 48-52% amino acid sequence identity with the other typical dehydrin proteins in plant cells. When the amino acid sequence of their proteins were aligned, several peptide motifs were well conserved, of which function has to be solved. Particularly the C-DH contains 15 additional amino acids at its N-terminus. Genomic Southern blot analysis using the coding region of C-DH showed that the C-DH consists of a single copy gene in Chinese cabbage genome. The C-DH mRNA, whose transcript size is 0.7 kb, was expressed with a tissue-specific manner. It was highly expressed in seed, flower buds and low expression as detected in root, stem or leaf tissues of Chinese cabbage. And the transcript level of C-DH was significantly induced by the treatment of plant hormone, abscisic acid and water-deficit conditions.

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Geographic homogeneity and high gene flow of the pear psylla, $Cacopsylla$ $pyricola$ (Hemiptera: Psyllidae), detected by mitochondrial COI gene and nuclear ribosomal internal transcribed spacer 2

  • Kang, Ah-Rang;Baek, Jee-Yeon;Lee, Sang-Hyun;Cho, Young-Sik;Kim, Wol-Soo;Han, Yeon-Soo;Kim, Ik-Soo
    • Animal cells and systems
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    • v.16 no.2
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    • pp.145-153
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    • 2012
  • The pear psylla, $Cacopsylla$ $pyricola$ (Hemiptera: Psyllidae), is a serious insect pest of commercial pear crops. The species, which resides on pear trees throughout its life cycle, is rapidly spreading in some regions of the world. The population genetic structure of the species collected from several pear orchards in Korea was studied to understand the nature of dispersal and field ecology of the species. The 658-bp region of mitochondrial COI gene and the 716-bp long complete internal transcribed spacer 2 (ITS2) of the nuclear ribosomal DNA were sequenced. Unlike other previously studied insect pests, the COI-based genetic diversity of the pear psylla was extremely low (maximum sequence divergence of 0.15%). This finding allowed us to conclude that the species may have been introduced in Korea relatively recently. ITS2 sequence-based analyses of phylogeny, population differentiation, gene flow, and hierarchical population structure all concordantly suggested that the pear psylla populations in Korea are neither genetically isolated nor hampered for gene flow. These genetic data are concordant with the dispersal of an overwintering winterform morph outside the non-pear habitat in the fall.

Cloning, Expression, and Characterization of DNA Polymerase from Hyperthermophilic Bacterium Aquifex pyrophilus

  • Choi, Jeong-Jin;Kwon, Suk-Tae
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.1022-1030
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    • 2004
  • The gene encoding Aquifex pyrophilus (Apy) DNA polymerase was cloned and sequenced. The Apy DNA polymerase gene consists of 1,725 bp coding for a protein with 574 amino acid residues. The deduced amino acid sequence of Apy DNA. polymerase showed a high sequence homology to Escherichia coli DNA polymerase I-like DNA polymerases. It was deduced by amino acid sequence alignment that Apy DNA polymerase, like the Klenow fragment, has only the two domains, the $3'{\rightarrow}5'$ exonuclease domain and the $5'{\rightarrow}3'$ polymerase domain, containing the characteristic motifs. The Apy DNA polymerase gene was expressed under the control of T7lac promoter on the expression vector pET-22b(+) in E. coli. The expressed enzyme was purified by heat treatment, and Cibacron blue 3GA and $UNO^{TM}$ Q column chromatographies. The optimum pH of the purified enzyme was 7.5, and the optimal concentrations of KCl and $Mg^{2+}$ were 20 mM and 3 mM, respectively. Apy DNA polymerase contained a double strand-dependent $3'{\rightarrow}5'$ proofreading exonuclease activity, but lacked any detectable $5'{\rightarrow}3'$ exonuclease activity, which is consistent with its amino acid sequence. The somewhat lower thermostability of Apy DNA polymerase than the growth temperature of A. pyrophilus was analyzed by the comparison of amino acid composition and pressure effect.

Cloning and Characterization of BTG-1 Gene from Pacific Oyster (Crassostrea gigas) (참굴(Crassostrea gigas)의 BTG1 유전자의 특성)

  • Chung, In Young;Oh, Jeong Hwan;Song, Young Hwan
    • Journal of Life Science
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    • v.27 no.4
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    • pp.398-407
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    • 2017
  • BTG 1 (B-cell translocation gene 1) gene was first identified as a translocation gene in a case of B-cell chronic lympocytic leukemia. BTG1 is a member of the BTG/TOB family with sharing a conserved N-terminal region, which shows anti-proliferation properties and is able to stimulate cell differentiation. In this study, we identified and characterized the pacific oyster Crassostrea gigas BTG1 (cg-BTG1) gene from the gill cDNA library by an Expressed Sequence Tag (EST) analysis and its nucleotide sequence was determined. The cg-BTG1 gene encodes a predicted protein of 182 amino acids with 57% 56% identities to its zebrafish and human counterparts, and is an intron-less gene, which was confirmed by PCR analysis of genomic DNA. Maximal homologies were shown in conserved Box A and B. The deduced amino acid sequence shares high identity with other BTG1 genes of human, rat, mouse and zebrafish. The phylogenic analysis and sequence comparison of cg-BTG1 with other BTG1 were found to be closely related to the BTG1 gene structure. In addition, the predicted promoter region and the different transcription-factor binding site like an activator protein-1 (AP-1) response element involved in negative regulation and serum response element (SRE) were able to be identified by the genomic DNA walking experiment. The quantitative real-time PCR analysis showed that the mRNA of cg-BTG1 gene was expressed in gill, heart, digestive gland, intestine, stomach and mantle. The cg-BTG1 gene was expressed mainly in heart and mantle.

Characterization of a Lactobacillus acidophilus Strain Isolated from Korean Infant Feces and Cloning of Surface Layer Protein Gene slp and Its Expression in Escherichia coli (유아 분변에서 분리한 Lactobacillus acidophilus의 특성 및 표면 단백질 유전자 클로닝과 대장균 내에서의 발현)

  • Park, Myeong-Soo;Ji, Geun-Eog;You, Kwan-Hee;Lee, Si-Kyung;Jeong, Won-Seok;Kim, Jin-Hyung;Jo, Myoung-Hwan;Kim, Soo-Young
    • Microbiology and Biotechnology Letters
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    • v.35 no.4
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    • pp.352-356
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    • 2007
  • A Lactobacillus sp. has been isolated from infant feces and characterized according to its physiological properties and identified as Lactobacillus acidophilus KLA1012. A gene coding surface layer protein (SLP) has been cloned and the sequence has been determined. The nucleotide sequence of slpA was 1,338 bp in size and was identical to that of L. acidophilus ATCC 4356 (100%). Amino acid sequence of SLP-A was deduced from the nucleotide sequence and it had signal sequence at N-terminal, consisting of positively charged amino acid mainly lysine. slpA was cloned and heterologously expressed in E. coli M15 and the 45.2 kDa surface-layer protein band was examined by SDS-PAGE and confirmed by Western blotting using polyclonal antibody against L. acidophilus KLA 1012 SLP-A protein.

Taxonomy of $Ganoderma$ $lucidum$ from Korea Based on rDNA and Partial ${\beta}$-Tubulin Gene Sequence Analysis

  • Park, Young-Jin;Kwon, O-Chul;Son, Eun-Suk;Yoon, Dae-Eun;Han, Woo-Ri-Ja-Rang;Yoo, Young-Bok;Lee, Chang-Soo
    • Mycobiology
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    • v.40 no.1
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    • pp.71-75
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    • 2012
  • In the present study, a phylogenetic analysis was undertaken based on the internal transcribed spacer (ITS) rDNA and partial ${\beta}$-tubulin gene sequence of the $Ganoderma$ species. The size of the ITS rDNA regions from different $Ganoderma$ species varied from 625 to 673 bp, and those of the partial ${\beta}$-tubulin gene sequence were 419 bp. Based on the results, a phylogenetic tree was prepared which revealed that Korean $Ganoderma$ $lucidum$ strains belong in a single group along with a $G.$ $lucidum$ strain from Bangladesh.

Intragenic Control of Expression of a Rice MADS Box Gene OsMADS1

  • Jeon, Jong-Seong;Lee, Sichul;An, Gynheung
    • Molecules and Cells
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    • v.26 no.5
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    • pp.474-480
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    • 2008
  • OsMADS1 is a rice MADS box gene necessary for floral development. To identify the key cis-regulatory regions for its expression, we utilized transgenic rice plants expressing GUS fusion constructs. Histochemical analysis revealed that the 5.7-kb OsMADS1 intragenic sequences, encompassing exon 1, intron 1, and a part of exon 2, together with the 1.9-kb 5' upstream promoter region, are required for the GUS expression pattern that coincides with flower-preferential expression of OsMADS1. In contrast, the 5' upstream promoter sequence lacking this intragenic region caused ectopic expression of the reporter gene in both vegetative and reproductive tissues. Notably, incorporation of the intragenic region into the CaMV35S promoter directed the GUS expression pattern similar to that of the endogenous spatial expression of OsMADS1 in flowers. In addition, our transient gene expression assay revealed that the large first intron following the CaMV35S minimal promoter enhances flower-preferential expression of GUS. These results suggest that the OsMADS1 intragenic sequence, largely intron 1, contains a key regulatory region(s) essential for expression.

Molecular Cloning and Characterization of the Rod Opsin Gene in Olive Flounder Paralichthys olivaceus

  • Kim, Jong-Myoung;Kim, Sung-Wan;Kim, Sung-Koo
    • Fisheries and Aquatic Sciences
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    • v.10 no.1
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    • pp.8-15
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    • 2007
  • Rhodopsin, a dim-light receptor, is a model system for the study of G protein-coupled receptors that transduce extracellular signals into cells. To study the molecular mechanisms of visual systems in fish, the rod opsin gene of olive flounder Paralichthys olivaceus was characterized. The full-length P. olivaceus opsin gene was obtained by PCR amplification of genomic DNA, as well as cDNA synthesis. A comparison of clones obtained from both methods indicated that the olive flounder rod opsin gene lacks introns. Sequence analysis of the opsin gene indicated that it contains a 1,056-bp open reading frame encoding 352 amino acids. The deduced amino acid sequence contains features of typical rod opsins, such as sites for Schiff's base formation (K296) and its counterion (E113), disulfide formation (C110 and C187), and palmitoylation (C322 and C323). An opsin sequence alignment showed the highest similarity between P. olivaceus and Solea solea (95.1%), followed by Hippoglossus hippoglossus (94.5%). An opsin phylogenetic tree revealed a close relationship between olive flounder and teleost rod opsins.

Transcriptional Regulation of the Gene Encoding ${\gamma}$-Glutamylcysteine Synthetase from the Fission Yeast Schizosaccharomyces pombe

  • Kim, Su-Jung;Kim, Hong-Gyum;Kim, Byung-Chul;Kim, Kyunghoon;Park, Eun-Hee;Lim, Chang-Jin
    • Journal of Microbiology
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    • v.42 no.3
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    • pp.233-238
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    • 2004
  • Transcriptional regulation of the Schizosaccharomyces pombe y-glutamylcysteine synthetase (GCS) gene was examined using the two GCS-lacZ fusion plasmids pUGCS101 and pUGCS102, which harbor 607 bp and 447 bp upstream regions, respectively. The negatively-acting sequence was located in the -607 - -447 bp upstream region of the GCS gene. The upstream sequence responsible for induction by menadione(MD) and L-buthionine-(S, R)-sulfoximine (BSO) resides in the -607 - -447 bp region, whereas the sequence which codes for nitric oxide induction is located within the -447 bp region, measured from the translational initiation point. Carbon source-dependent regulation of the GCS gene appeared to be dependent on the nucleotide sequence within -447 bp region. The transcription factor Papl is involved in the induction of the GCS gene by MD and BSO, but not by nitric oxide. Induction of the GCS gene occurring due to low glucose concentration does not depend on the presence of Pap1. These data imply that induction by MD and BSO may be mediated by the Pap1 binding site, probably located in the -607 - -447 region, and also that the nitric oxide-mediated regulation of the S. pombe GCS gene may share a similar mechanism with its carbon-dependent induction.