• 제목/요약/키워드: ITS gene sequence

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Random Sequence Analysis of the Genomic DNA of Methanopyrus kandleri and Molecular Cloning of the Gene Encoding a Homologue of the Catalytic Subunit of Carbon Monoxide Dehydrogenase

  • Shin, Hyun-Seock;Ryu, Jae-Ryeon;Han, Ye-Sun;Choi, Yong-Jin;Yu, Yeon-Gyu
    • Journal of Microbiology and Biotechnology
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    • 제9권4호
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    • pp.404-413
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    • 1999
  • Methanopyrus kandleri is a hyperthermophilic methanogen that represents one of the most heat-resistant organisms: the maximum growth temperature of M. kandleri is $110^{\circ}C$. A random sequence analysis of the genomic DNA of M. kandleri has been performed to obtain genomic information. More than 200 unique sequence tags were obtained and compared with the sequences in the GenBank and PIR databases. About 30% of the analyzed tags showed strong sequence similarity to previously identified genes involved in various cellular processes such as biosynthesis, transport, methanogenesis, or metabolism. When statistics relating to the frequency of codons were examined, the sequenced open reading frames showed highly biased codon usage and a high content of charged amino acids. Among the identified genes, a homologue of the catalytic subunit of carbon monoxide dehydrogenase (CODH) that reduces $CO_2$ to CO was cloned and sequenced in order to examine its detailed gene structure. The cloned gene includes consensus promoters. The amino acid sequence of the cloned gene shows a strong homology with the CODH genes from methanogenic Archaea, especially in the presumed binding sites for Fe-S centers.

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Cloning of a Laccase Gene Fragment from Ganoderma lucidum (영지버섯 Laccase 유전자의 구리결합부위 I과 IV사이 지역의 클로닝)

  • 조지현;최형태;김경훈
    • Korean Journal of Microbiology
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    • 제36권3호
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    • pp.192-195
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    • 2000
  • Degenerate primevs corresponding to the consensus sequences of the copper-binding regions in the N- and Cterminal domains of fungal laccases were used to isolate laccase gene-specific sequences froin a white rot rungus Ganodern~a lucidrm w-hich has been known to strengthen the imnnne system. A 1.6 Kbp fragment was amplified by PCR and its base sequence was detenuiued. Locating seven iutrous within the base sequence, we could deduce its amino acid sequence. The nucleotide sequence witl~out introlls was 47Y0 identical to that of lee1 gene of Pametes wllosa; lhe identity in amino acid sequences of the two was 7994 The deduced amino acid seqoence was also sunilar to those of Coriolus versicolo~ kc3 (79%); Co~,iolz~s hirsutus phenolouiduse (78%), Trainetes vel.srcoloi. lccl (77%), Trametes ~!i/Iosa Ice2 (77%) and Trametes vemicolor kc4 (66%).

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Nucleotide Sequence and Inducibility Analysis of Chloramphenicol Acetyltransferase Gene from Staphylococcus aureus R-plasmid pSBK203 (Staphylococcus aureus에서 분리된 R-plasmid pSBK203상의 chloramphenicol acetyltransferase 인자의 염기서열 및 유발성 분석)

  • 권동현;변우현
    • Korean Journal of Microbiology
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    • 제27권3호
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    • pp.194-200
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    • 1989
  • The nucleotide sequence of inducible chloramphenicol acetyl-transferase(CAT) gene isolated from a small plasmid pSBK203 of Staphylococcus aureus was determined. The base sequence shows that structural gene of pSBK203-CAT encodes a protein of 213 amino acids and has a leader region which encodes a short polypeptide of 9 amino-acids in its upstream. vertical bar /sup 35/S vertical bar-Methionine labelled CAT gene product in minicell showed almost same mobility with pC194-CAT of which molecular weight is 24Kdal on polyacrylamide gel electrophoresis. Predicted amino acid sequence of pSBK203-CAT has revealed a high degree of homology with the CATs of pC194 and pC221 than those of cat-86, Tn9 and proteus mirabilis PM13.

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Nucleotide sequence analysis of the 5S ribosomal RNA gene of the mushroom tricholoma matsutake

  • Hwang, Seon-Kap;Kim, Jong-Guk
    • Journal of Microbiology
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    • 제33권2호
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    • pp.136-141
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    • 1995
  • From a cluster of structural rRNA genes which has previsouly been cloned (Hwang and Kim, in submission; J. Microbiol. Biotechnol.), a 1.0-kb Eco RI fragment of DNA which shows significant homology to the 25S and rRNA s of Tricholoma matsutake was used for sequence analysis. Nucleotide sequence was bidirectionally determined using delection series of the DNA fragment. Comparing the resultant 1016-base sequence with sequences in the database, both the 3'end of 25S-rRNA gene and 5S rRNA gene were searched. The 5S rRNA gene is 118-bp in length and is located 158-bp downstream of 3'end of the 25S rRNA gene. IGSI and IGS2 (partial) sequences are also contained in the fragment. Multiple alignment of the 5S rRNA sequences was carried out with 5S rRNA sequences from some members of the subdivision Basidiomycotina obtained from the database. Polygenetic analysis with distance matrix established by Kimura's 2-parameter method and phylogenetic tree by UPGMA method proposed that T. matsutake is closely related to efibulobasidium allbescens. Secondary structure of 5S rRNA was also hypothesized to show similar topology with its generally accepted eukaryotic counterpart.

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Cloning and Expression of the Aminopeptidase Gene from the Bacillus lichenformis In Bacillus subtilis

  • Kim, Jin-Sook;Lee, In-Soo;Lee, Seung-Won;Lee, Young-Phil;Jung, Chul-Ho;Kim, Hyung-Cheol;Choi, Soon-Yong
    • Journal of Microbiology and Biotechnology
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    • 제12권5호
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    • pp.773-779
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    • 2002
  • A gene (hap) encoding aminopeptidase from the chromosomal DNA of Bacillus licheniformis was cloned. The gene is 1,347 bp long and encodes a 449 amino acid preproprotein with a major mature region of 401 amino acids (calculated molecular mass 43,241 Da). N-Terminal sequence of the purified protein revealed a potential presence of N-terminal propeptide. The deduced primary amino acid sequence and the mass analysis of the purified protein suggested that a C-terminal peptide YSSVAQ was also cleaved off by a possible endogeneous protease. Tho amino acid sequence displayed 58% identity with that of the aminopeptidase from alkaliphilic Bacillus halodurans. This bacterial enzyme was overexpressed in recombinant Escherichia coli and Bacillus subtilis cells. Clones containing the intact hap gene, including its own promoter and signal sequence, gave rise to the synthesis of extracellular and thrmostable enzyme by B. subtilis transformants. The secreted protein exhibited the same biochemical properties and the similar apparent molecular mass as the B. lichenzyormis original enzyme.

Nucleotide Sequence of a Bacteriolytic Enzyme Gene from Alkalophilic Bacillus sp.

  • Jung, Myeong-Ho;Ohk, Seung-Ho;Yum, Do-Young;Kong, In-Soo;Bai, Dong-Hoon
    • Journal of Microbiology and Biotechnology
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    • 제3권2호
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    • pp.73-77
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    • 1993
  • The nucleotide sequence of Bacillus sp. bacteriolytic enzyme gene, lytP and its flanking regions were determined. A unique open reading frame for a protein of Mw. 27, 000, and a putative terminator sequence, were found behind a concensus ribosome binding site located 8 nt upstream from ATG start codon. The primary amino acid sequence deduced from nucleotide sequence revealed a putative protein of 255 amino acid residues with an Mw. of 27, 420. No significant homology could be found between the amino acid sequence of Bacillus sp. bacteriolytic enzyme and that of other cell wall hydrolases.

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The Base Sequence of ITS and Genetic Variation in Sarcodon Aspratus (능이버섯의 ITS염기서열과 유전적 변이)

  • Kim Jong Bong
    • Journal of Life Science
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    • 제14권6호
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    • pp.963-966
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    • 2004
  • The sequence of ITS (partial 16S ribosomal DNA, complete ITS1, 5.8S ribosomal DNA and ITS2, and partial 28S ribosomal DNA) was analysed by PCR and autosequencing in Sarcodon aspratus. The ITS lenght of S. aspratus was 716 base pair. As this sequence compared with other reports on S. aspratus (ace No AF335110), the sequence variation based on nucleotide deletion and substitution was $1.8\%$. This nucleotide variation rate in same species was very higher than in other species. Also, the sequence varitation rates between this S. aspratus and S. imbricatus, and S. squamus were $8\%\;and\;10\%$, respectively. This results suggested that the high sequence variation of S. aspratus might be caused specific host and inhabitat environment which limited gene flow.

Nucleotide Sequence of the Penicillin G Acylase Gene from Bacillus megaterium and Characteristics of the Enzyme (Bacillus megaterium에서 발견된 Penicillin G Acylse 유전자의 염기서열과 그 효소의 특성)

  • Gang, Ju-Hyeon;Kim, Seong-Jae;Park, Yong-Chjun;Hwang, Young;Yoo, Ook-Joon;Kim, Young-Chang
    • Korean Journal of Microbiology
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    • 제32권3호
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    • pp.215-221
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    • 1994
  • The complete nucleotide sequence of the cloned pga gene encoding the penicillin G acylase of Bacillus megaterium ATCC 14945 and its 5'- and 3'-flanking regions was determined. The sequence revealed only one large open reading frame (2,406 hp) of the penicillin G acylase (pga) gene. Upstream from ATG of the pga gene, there was a putative ribosome binding site, Shine-Dalgarno sequence. The promoter-like structure, - 10 and - 35 sequences, was also found. Following the stop codon, TAG, a structure reminiscent of the E. coli rho-independent transcription terminator was present. The amino acid sequence was deduced from the nucleotide sequence. The molecular mass of the polypeptide was 91,983 Da. There was a potential signal sequence in its amino-terminal region. A comparison of its deduced amino acid sequence with other characterized penicillin G acylases and the result of SDS-polyacrylamide gel electrophoresis of the purified enzyme showed that a precursor polypeptide of 92 kDa was processed into two dissimilar ${\alpha}$ and ${\beta}$-subunits of 25 and 61 kDa.

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NaCl-dependent Amylase Gene From Badillus circulans F-2 Its Nucleotide Sequence (Bacillus circulans F-2의 NaCl 의존성 amylase 유전자의 DNA 염기배열 결정)

  • 김철호;권석태;타니구치하지메;마루야마요시하루
    • Microbiology and Biotechnology Letters
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    • 제18권3호
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    • pp.309-316
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    • 1990
  • The sequence of a 1795 bp restriction fragment containing the B. circulans F-2 gene for NaC1- dependent $\alpha$-amylase (CI-amylase) is reported. The probable coding region of the gene is 1005 base pairs (335 amino acida) long. The NaC1-dependent $\alpha$-amylase (el-amy) sequence shows an open reading frame (ORF) with the translated molecular weight of about 38, 006, which correspond to a molecular weight of about 35, 000 (Mi). The gene is preceded by the sequence resembling promoter for the vegetative B, subtitis RNA polymerases. These are followed by the sequences resembling a B. subtilis ribosome binding site 5 nucleotides before the first codon of the gene. Homologous regions with other amylases were found. The N-terminal sequences of the mature proteins expressed in E. eoli were identical to the N-terminal sequences which are anaIysed.

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Intron sequence diversity of the asian cavity-nesting honey bee, Apis cerana (Hymenoptera: Apidae)

  • Wang, Ah Rha;Jeong, Su Yeon;Jeong, Jun Seong;Kim, Seong Ryul;Choi, Yong Soo;Kim, Iksoo
    • International Journal of Industrial Entomology and Biomaterials
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    • 제31권2호
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    • pp.62-69
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    • 2015
  • The Asian cavity-nesting honeybee, Apis cerana (Hymenoptera: Apidae), has been extensively studied for its biogeography and genetic diversity, but the molecules utilized in past studies were mainly ~90 bp long mitochondrial non-coding sequences, located between $tRNA^{Leu}$ and COII. Thus, additional molecular markers may enrich our understanding of the biogeography and genetic diversity of this valuable bee species. In this study, we reviewed the public genome database to find introns of cDNA sequences, with the assumption that these introns may have less evolutionary constraints. The six introns selected were subjected to preliminary tests. Thereafter, two introns, titled White gene and MRJP9 gene, were selected. Sequencing of 552 clones from 184 individual bees showed a total of 222 and 141 sequence types in the White gene and MRJP9 gene introns, respectively. The sequence divergence ranged from 0.6% to 7.9% and from 0.26% to 17.6% in the White gene and the MRJP9 introns, respectively, indicating higher sequence divergence in both introns. Analysis of population genetic diversity for 16 populations originating from Korea, China, Vietnam, and Thailand shows that nucleotide diversity (π) ranges from 0.003117 to 0.025837 and from 0.016541 to 0.052468 in the White gene and MRJP9 introns, respectively. The highest π was found in a Vietnamese population for both intron sequences, whereas the nine Korean populations showed moderate to low sequence divergence. Considering the variability and diversity, these intron sequences can be useful as non-mitochondrial DNA-based molecular markers for future studies of population genetics.