• 제목/요약/키워드: cloning homology

검색결과 262건 처리시간 0.026초

Cloning and Characterization of the psbEF Gene Encoding Cytochrome b-559 of the Panax ginseng Photosystem II Reaction Center

  • Lee, Won-Kyu;Park, Dae-Sung;Tae, Gun-Sik
    • BMB Reports
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    • 제32권2호
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    • pp.189-195
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    • 1999
  • From the Panax ginseng chloroplast, the psbE and psbF genes, encoding the $\alpha$- and $\beta$-subunits of cytochrome b-559 of the photosystem II reaction center, respectively, were cloned and characterized. The psbE and psbF genes were composed of 252 and 117 nucleotides, respectively. The deduced amino acid sequence of the $\alpha$-subunits showed 95%, 93%, and 91% homology to monocots, dicots, and liverwort, respectively, whereas the $\beta$-subunits showed approximately 98% to 95% homology to the same species. Southern blot analysis revealed that a single copy of the psbEF gene exists in the chloroplast plastid. Northern blot analysis indicated that the psbE and psbF genes are cotranscribed as a polycistron.

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Cloning and Sequenece Analysis of the hpa D Gene Responsible for Homoprotocatechuate 2, 3-Dioxgenae from Pseudomonas sp. DJ-12

  • Lee, Sang-Maha;Chae, Jong-Chan;Kim, Young-Soo;Kim, Chi-Kyung
    • Journal of Microbiology
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    • 제39권4호
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    • pp.334-337
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    • 2001
  • The degradative pathway of homoprotocatechuate (HPC) is the bacterial routhe wherby 3,4-dihydrox-yphenylactic acid is catabolized to pyruvate and succinate by a series of sequential reactions . The HPC is catalzed by homoprotocatechuate 2, 3-dioxygenase(HPC-2,3O) to from 5-carboxymethy1-2-hydroxy-muco semialdehyde. In this study, the hha D gene encoding. HPC, 2, 3O was Cloned from the chromo-somal DNA of Pseudomonas sp. DJ-12 and its nucleotide sequence was analyzed. The open reding frame of hpaD gene was found to be composed of 864 nucleotide pairs and to encode a poypetide with 287 amino acide residues. The deduced amino acid sequence of the HPC-2,3O from Pseudomonas. sp. DJ-12 exhibited 60~64% homology with those of the corresponding enzymes from E. coli. Salmonella enterica, and Klebsiella pneumoniae.

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생쥐 섬 유아세포에서 70 kDa 고온충격 단백질의 CDNA 클로닝과 염기서열 분석 (Isolation and Characterization of a CDNA Encoding a Protein Homologous to the Mouse 70 kDa Heat Shock Protein)

  • 김창환;정선미최준호
    • 한국동물학회지
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    • 제35권2호
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    • pp.203-210
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    • 1992
  • Hsp70, a 70 kDa protein, is the maior protein expressed when cells are heat-shocked. A cDNA library from mouse ID13 cells was screened with the human hsp70 gene as a probe, and a positive clone was obtained. The positive clone was subcloned into puc19 and the precise restriction was obtained. The CDNA was sequenced by the Sanger's dideoxv termination method. Single open reading frame that codes for a protein of 70 kDa was found. The DNA sequence of the cloned mouse DNA shows great homology (66-90%) with other mouse hsp70 genes and somewhat less homology (50",) with E. coli hsp70 gene (dnak). With the exception of one amino acid, the protein sequence deduced from the CDNA is identical to the mouse that shock cognate protein 70 (hsc70) that is constitutivelv expressed at normal temperature. The result suggests that the cloned CDNA encodes a hsc70 family rather than a heatinducible family.mily.

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토마토에서 분리된 담배 모자이크 바이러스 외피단백질 유전자의 cDNA 클로닝 및 염기서열 분석 (Complementary DNA Cloning and Nucleotide Sequence Analysis of Coat Protein Gene from TMV Tomato Strain)

  • 이청호;이영기;강신웅;박은경
    • 한국연초학회지
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    • 제18권2호
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    • pp.101-106
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    • 1996
  • Tobacco mosaic virus (TMV) tomato strain was isolated from tomato "Seo-Kwang" in Korea. The virion was purified by density gradient centrifugation, and total viral RNA was isolated from the purified particles. Coat protein (CP) cDNA of the virus was synthesized by RT-PCR, and the purified cDNA fragment was subcloned to pBluescript II SK-. The analysis of nucleotide sequence showed that this cDNA was 693 nucleotides long from the insert of clone p1571 and p1572 which contain complete codons of the viral coat protein gene (474 nucleotides) and 3' untranslated region. The nucleotides of coat protein encoding cDNA of the strain were 6 nucleotides less than that of TMV common strain isolated from tobacco plant in Korea. The CP gene showed 70% maximum homology with that of the common strain in the nucleotide level and 86% maximum homology in amino acid level.cid level.

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Staphylococcus aureus DH1에서 분리된 R-plasmid pSBK203의 복제 개시 유전자(rep) 분리 및 염기서열 결정 (Cloning and Base Sequence Determination of Replication Initiation Gene (rep) Isolated from Staphylococcus aureus DH1 R-plasmid pSBK203)

  • Park, Seung-Moon;Kwon, Dong-Hyun;Byeon, Woo-Hyeon
    • 미생물학회지
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    • 제31권1호
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    • pp.44-47
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    • 1993
  • A replication initiation gene was identified and its nucleotide sequence has been determined from a 3.8 kb, chloramphenicol acethyltransferase conferring R-plasmid pSBK203 of Staphylococcus aures. Location of the replication related region of pSBK 203 was determined by interuption with pUC 119 at XBaI and MspI sites which resulted in inactivation of replication in Bacilius subtilis. Base sequence of this region revealed on open reading frame of 942 base pairs, which encoded a 314 amino acid protein. Base sequence homology with other rep of pT181 family plasmids such as pT181, pC221, pC223, pS194, pU112, and pCW7 was ranged from 78% to 97% and the predicted amino acid sequence homology was from 72% to 95%.

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Cloning of the $Glu-tRNA^{Gln}$ Amidotransferase (gatCAB) Gene from Staphylococcus aureus

  • Namgoong, Suk;Hong, Kwang-Won;Lee, Se-Yong
    • Journal of Microbiology and Biotechnology
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    • 제11권2호
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    • pp.251-258
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    • 2001
  • In Gram(+) bacteria and organelles in higher eukarotes, $Gln-tRNA^{Gln}$ utilized for protein biosynthesis is formed by a tRNA-dependent amino acid transformation using mischarged $Gln-tRNA^{Gln}$ as the intermediate. In this study, the gatCAB gene encoding $Gln-tRNA^{Gln}$ amidotransferase (Glu-AdT) of Staphylococcus aureus was cloned and its nucleotide sequence wa determined. The S. aureus gatCAB gene was organized in an operon structure consisting of three open reading frames (gatC, gatA, and gatB), similar to that of Bacillus subtilis. The gene sequences for the A and B subunits of$Gln-tRNA^{Gln}$ amidotransferase showed significant homology (77 and 87% homology with amino acid sequence) with the gatA and gatB genes of B. subtilis, yet the C subunit (gatC) showed a relatively lowe homology with the B. subtilis gatC gene and other orthologues. The cloned S. aureus <$Gln-tRNA^{Gln}$ amidotransferase gene was highly expressed in Escherichia coli, and the resulting crude enzyme could convert misacylated <$Gln-tRNA^{Gln}$ into $Gln-tRNA^{Gln}$ in vitro.

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Homology Modeling and In Vitro Analysis for Characterization of Streptomyces peucetius CYP157C4

  • Rimal, Hemraj;Yu, Sang-Cheol;Jang, Jong Hwa;Oh, Tae-Jin
    • Journal of Microbiology and Biotechnology
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    • 제25권9호
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    • pp.1417-1424
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    • 2015
  • In this study, we tried to characterize Streptomyces peucetius CYP157C4 with homology modeling using three cytochrome P450 (CYP) structures (CYP157C1, CYP164A2, and CYP107L1), having discovered that CYP157C4 lacks the ExxR motif that was considered invariant in all CYPs. We used Discovery Studio 3.5 to build our model after first assessing the stereochemical quality and side-chain environment, and a 7-ethoxycoumarin substrate was docked into the final model. The model-substrate complex allowed us to identify functionally important residues and validate the active-site architecture. We found a distance of 4.56 Å between the 7-ethoxycoumarin and the active site of the heme, and cloning and an in vitro assay of the CYP157C4 showed the dealkylation of the substrate. Since the details regarding this group of CYP structures are still unknown, the findings of this study may provide elucidation to assist with future efforts to find a legitimate substrate.

러시아범안열원충(Theileria sergenti) 국내 분리주의 33 kDa piroplasm protein 유전자 크로닝 및 염기서열 (Cloning and sequencing of p33 in a Korean isolate of Theileria sergenti)

  • 강승원;최은진;권창희
    • Parasites, Hosts and Diseases
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    • 제35권2호
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    • pp.105-110
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    • 1997
  • T. sergenti 국내 분리주의 면역항원인 33 kDa의 piroplasm surface protein 유전자를 크로닝하였다. 크로닝된 T. sergenti의 33 kDa에 해당하는 유전자의 염기서열을 분석한 결과 총 869 bps의 염기와 283개의 아미노산을 확인하였다. 또 이들 분석결과를 일본주의 염기서열 및 아미노산 조성과 비교 분석하였던 바 각각 99.4. 98.9%의 homology를 나타내었으므로 두 주간의 p33 유전자는 거의 일치하는 것으로 판명되었다.

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해양미생물 Streptomyces sp. M3로부터 alginate lyase의 클로닝 및 발현 (Cloning and Expression of Alginate Lyase from a Marine Bacterium, Streptomyces sp. M3)

  • 김희숙
    • 생명과학회지
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    • 제19권11호
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    • pp.1522-1528
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    • 2009
  • 알긴산을 분해하기 위하여 갈조류로부터 분해활성이 있는 해양미생물을 분리하였다. 분리된 균주의 16S ribosomal DNA를 분석한 결과 이전에 보고했던 ALG-5 균주와 비슷한 Streptomyces sp.에 속하는 것으로 나타났다. 상동성이 있는 염기서열로 고안한 특이적인 primer로 PCR을 행함로서 Streptomyces sp. M3의 새로운 alginate lyase 유전자를 클로닝하였다. M3 alginate lyase의 예상 아미노산 서열에는 N-terminal 영역에 YXRSELREM 서열과 C-terimnal 영역에 YFKAGXYXQ 서열이 보존되어 있었다. M3 alginate lyase 단백질의 homology model은 Corynebacterium sp. ALY-1으로부터 얻은 단백질인 alyPG와 같이 $\beta$-jelly roll fold를 main domain으로 가지고 있음이 나타났다. M3 alginate lyase 유전자를 가지는 재조합 E. coli의 세포균질액은 polymannuronate block보다는 polyguluronate block에 대하여 높은 분해력을 가지고 있었다. 아미노산 서열 다중정열 및 homology modeling으로부터 얻은 결과는 M3 alginate lyase가 Family PL-7으로 분류될 수 있음을 말해 준다.

ISOLATION AND CHERACTERIZATION OF ACTINIDIN GENE FROM CHINESE WILD KIWI FRUIT

  • Lee, Nam-Keun;Hahm, Young-Tae
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 춘계학술발표대회
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    • pp.527-530
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    • 2000
  • 중국산 야생키위로 total RNA를 추출하고 RT-PCR를 실시해서 증폭된 1.2kb fragment를 얻어 pGEM-T Easy에 cloning한 후 염기서열과 아미노산 서열을 비교 분석한 결과 actinidin gene인 것으로 분석되었다.

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