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

  • 조지현;최형태;김경훈
    • 미생물학회지
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    • 제36권3호
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    • pp.192-195
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    • 2000
  • 백색부후균류 laccase의 아미노 말단과 카복시 말단에 잘 보존된 구리결합부위를 암 호화하는 DNA 염기서열과 상보적인 primer를 이용하여 백색부후균의 일종인 영지버섯 Gandoderma lucidum에서 laccase 유전자 단편을 분리하였다. PCR 로 증폭하여 클로닝한 1.6 Kb DNA 절편의 염기 서열을 결정하여 분석하였다. 이 DNA에는 7개의 인트론이 존재 하였으며 엑손의 염기서열과 이로부터 추정된 아미노산 서열은 Tranmetes villosa laccase(lccl)와 각각 47%, 79% 동일하였다. 기타 다른 백색부후균류의 laccase 아미노산 서 열과는 66~78% 동일하였다.

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

  • 권동현;변우현
    • 미생물학회지
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    • 제27권3호
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    • pp.194-200
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    • 1989
  • S. aureus에서 분리된 plasmid pSBK203 상의 CAT 유전자 염기서열을 결정하였으며 유발성 발현현상이 확인되었다. 염기서열 결과에 의해 예측된 단백질의 아미노산 서열 분석결고 pC221-CAT 와는 78%의 가장 높은 상동성을 나타냈으며 pC194-CAT와는 55%, 그람음성균 유래의 CAT 중 하나인 Tn9-CATdhkss 38%의 상동성을 각각 보여주고 있었다.

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

  • 김종봉
    • 생명과학회지
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    • 제14권6호
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    • pp.963-966
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    • 2004
  • 능이버섯의 16S ribosomal DNA일부분, IT1, 5.85 ribosomal DNA, ITS2의 전부분, 28S ribosomal DNA 일부분이 포함된 ITS영역의 염기서열을 분석하였다. 이 부분은 716개의 염기쌍으로 구성되었다. 이를 Sarcadon속에 속하는 종들과 비교분석한 결과 같은 능이버섯의 ITS에 관한 다른 분석결과 염기치환 및 결실을 근거로 하였을 경우 $1.8\%$의 차이를 나타내었다. 또한 S. imbricatus와는 $1.8\%$, S. sequamous와는 $10\%$차이를 나타내었다. 이는 능이버섯이 숙주, 서식지 환경 등의 특수성 때문에 자연상태에서 유전자교류가 일어나지 않기 때문인 것으로 생각된다.

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

  • 강주현;김성재;박용춘;황영;유욱준;김영창
    • 미생물학회지
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    • 제32권3호
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    • pp.215-221
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    • 1994
  • Bacillus megaterium ATCC 14945의 penicillin G acylase 유전자의 염기배열을 결정하였다. 이 유전자에는 2,406 염기쌍으로 이루어진 하나의 open reading frame이 존재하는데, 개시코돈의 5' 위쪽에서 Shine-Dalgarno 배열과 promoter로 여겨지는 부분을 발견하였으며, 종결코돈의 3' 아래쪽에서 rho-independent한 전사종결체와 dby사한 구조를 발견하였다. 염기배열로부터 폴리펩티드의 아미노산 배열을 유추하였다. 이 폴리펩티드의 분자량은 91,983 Da이었으며, 아미노 말단 부이에 signal sequence가 존재하였다. 이 아미노산 배열을 여러 다른 penicillin G acylase의 아미노산 배열과 비교하고 분리 정제한 효소를 SDS-polyacrylamide gel 전기영동으로 분석한 결과로부터 이 효소는 92kDa의 전구체로 해독된 후 processing 과정을 거쳐 각각 25kDa과 61kDa의 ${\alpha}$-, ${\beta}$-단위체로 구성됨을 알 수 있었다.

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

  • 김철호;권석태;타니구치하지메;마루야마요시하루
    • 한국미생물·생명공학회지
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    • 제18권3호
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    • pp.309-316
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    • 1990
  • Bacillus circulans F-2의 생산하는 NaCl 의존성 amylase(NaCl-dependent amylase) 유전자를 함유하는 1795bp의 DNA 염기배열을 결정하였다. 본 유전자의 ORF는 총염기수 1005bp(335 아미노산)로 구성되며, 분자량 38,006의 amylase의 분자량 약 35,000과 일치하였다. 본 유전자의 상류영역(upstream region)에는 고초균(Bacillus subtiis)의 전형적인 전사발현영역(transcriptional region)과 상보적인 DNA역역이 존재하였다. 성숙단백질의 N-말단측 아미노산 배열은 Ala-Ser-Lys-Val-Gly이며, 분비에 필요한 20개의 signal 아미노산 배열을 갖는 전형적인 분비 단백질임이 확인 되었다. 한편 다른 amylase들과 비교결과, smylase 활성발현과 밀접히 관련되 있는 4개 부위의 상보성영역(homologous region)을 가지고 있었다.

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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.