• 제목/요약/키워드: 유전자클로닝

검색결과 499건 처리시간 0.033초

돼지 콜레라 바이러스 E2 유전자의 클로닝 및 염기서열분석 (Cloning and Sequence Analysis of Hog Cholera Virus(HCV) E2 Gene)

  • 이영기;강신웅;김선원;박성원;이종철;이청호
    • 한국연초학회지
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    • 제23권2호
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    • pp.103-108
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    • 2001
  • Hog cholera virus(HCV) was purified from virus infected Bovine kidney cells. From this virus, total protein was analyzed by SDS-PAGE gel electrophoresis and about 55 kDa band of E2 envelope protein was detected. The viral RNA was purified and E2 cDNA was amplified by RT-PCR. E2 cDNA fragment was cloned to PCRII-TOPO cloning vector and named pE2. The analysis of nucleotide sequence showed that this E2 cDNA fragment inserted into pE2 was 1191 nucleotides long and coded 397 amino acids.

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Pseudomonas putida의 Catechol 2,3-dioxygenase 유전자의 클로닝 (Cloning of Catechol 2,3-dioxygenase Gene from Pseudomonas putida)

  • 김영수;최봉수;민경락;김치경
    • 미생물학회지
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    • 제29권3호
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    • pp.155-159
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    • 1991
  • Four strains of Pseudomonas putida (NAH), Pseudomonas sp.(TOL), Achromobacter xylosoxidans, and Alcaligenes sp. were compared with their degradative capability of aromatic compounds. All of the bacterial strains were utilized catechol as a sole carbon source for growth, but signigicantly different in degradative properties for 5 other aromatic compounds. Catechol 2, 3-dioxygenase gene from P. putida (NAH) has been cloned and expressed in E. coli. The DNA clone designated pCNU101 contains NAH-derived 6 Kb insert and its physical map was characterized. A subclone (pCNU106) for the catechol dioxygenase gene in pCNU101 contained 2.0kb-DNA insery fragmented by HpaI and ClaI.

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비름에서 ATP 가수분해효소와 상동성을 가지는 유전자의 클로닝 (Molecular Cloning and Nucleotide Sequence of Amaranthus viridis Homologue of the H -Transporting ATPase Gene)

  • 한규웅
    • 생명과학회지
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    • 제6권1호
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    • pp.1-5
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    • 1996
  • Using differential hybridization, a cDNA clone was isolated fortuitously from Amaranthus viridis and sequenced. This nucleotide sequence exhibited 55.1% identity with vma6 which encodes the 36-kD subunit of the vacuolar proton transporting ATPase in Saccharmoyces cerevisiae. The predicted open reading frame encodes a protein of 221 amino acid sequence with a calculated molecular weight of 25,452 and reveals high levels of similarity with subunit D polypeptide of vacuolar H -ATP(e.g., 48.5, 52.1 and 49.3% identity to the vacuolar 36-kD chain of yeast, vacuolar 32-kD polypeptide IV of human and vacuolar 28-kD protein of bovine chromaffin granules, respectively). The hydropathy index computation revealed that this predicted protein is a peripheral protein. These results indicated that the predicted protein may play a sturctural role in the vaculor H -ATPase as does gamma subunit in V-type ATPase.

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완두콩(Pisum sativum)에서 Ribulose-1,5-Bisphosphate Carboxylase Small Subunit 유전자의 cDNA 클로닝과 광유도성 발현에 관한 연구 (Cloning of cDNA Encoding the Precursor to the Small Subunit of Ribulose-1,5-Bisphosphate Carboxylase in Pea 9Pisum sativum))

  • 김한집
    • Journal of Plant Biology
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    • 제32권1호
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    • pp.33-40
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    • 1989
  • Polysomal polyadenylated mRNAs which were purified from pea leaves were fractionated by sucrose grandient sedimentation. Fractions corresponding to the peak at 11.5S were found to contain mostly mRNA encoding the precursor polypeptide to the small subunit of ribulose bisphosphate carboxylase (rbcS) by in vitro translation in wheat germ extract. Double-stranded cDNA which was synthesized from the 11.5S mRNA was cloned into Hind III site of plasmid pBR 325. A cDNA clone, H24, was identified to code for rbcS. In vitro translation product of the hybridization-selected mRNA was molecular weight 20,000, presumably the precursor of rbcS. The nucleotide sequences of the H24 showed almost complete homology with the sequences encoding the transit peptide of the rbcS-3A gene which was reported by Fluhr et al.(1986).

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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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벼 렉틴 유전자의 클로닝 및 대장균에서의 발현 (Molecular Cloning and Expression of Rice Lectin in Escherichia coli)

  • 홍성관;전상훈;김하형;공광훈
    • 약학회지
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    • 제46권4호
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    • pp.270-275
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    • 2002
  • The lectin gene from rice was amplified by the polymerase chain reaction. The amplified DNA was inserted into the expression vector pET26b and expressed it as a fusion protein with polyhistidine sequences in Escherichia coli. The recombinant protein was produced by induction with 0.4 mM isopropyl-$\beta$-D-thiogalactopyranoside at 37$^{\circ}C$ and purified by an immobilized metal affinity chromatography. The recombinant protein was found to have lectin activity by the hemagglutination inhibition assay. The hemagglutination activity of the recombinant protein was optimal at pH 4.0-7.0 and was dependent on $Ca^{2+}$ and Mn$^{2+}$.2+/.

Bacillus licheniformis NBL420 유래의 Xylanase 유전자의 클로닝과 특성 검토 (Cloning and Characterization of Xylanase Gene from Bacillus licheniformis NBL420)

  • 홍인표;최신건
    • 산업기술연구
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    • 제29권A호
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    • pp.169-176
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    • 2009
  • The gene encoding endoxylanase (xylS) was isolated from a genomic library of Bacillus licheniformis NBL420. Two positive clones, which harbor 1.5 kb and 0.8 kb inserts respectively, were screened on RBB dyed-xylan plates and the recombinant plasmids were named as pBX3 and pBX5. The nucleotide sequencings of two inserts revealed the existence of common 639 bp of open reading frame which encode 232 amino acids. The xylS gene was successfully subcloned into pET22b(+) vector and overexpressed. Enzymatic properties including optimum pH, optimum temp, thermostability and pH stability were investigated. Activity staining of XylS was identical with that of original Bacillus licheniformis NBL420.

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Lactobacillus casei의 phospho-$\beta$-galactosidase 유전자의 대장균내 분자클로닝 (Molecular cloning of phospho-$\beta$-galactosidase gene of lactobacillus casei in escherichia coli)

  • 문경희;박정희;최순영;이유미;김태한;김연수;민경희
    • 미생물학회지
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    • 제27권3호
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    • pp.188-193
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    • 1989
  • Gene for lactose catabolism in Lactobacillus casei SW-M1 was encoded by a 60Kb metabolic plasmid. A derivative of only 10kb, pPlac 15 of recombinant plasmid, was constructed by introducing into pBR322 and was cloned into E. coli using restriction endonuclease Pst I. A 10kb insery DNA in plasmid pBR322 was identified as a gene encoded phospho-$\beta$-galactosidase by the determination of enzyme activity. Phospho-$\beta$-galactosidase was apparently expressed in E. coli. The enzyme activities of cell-free extract from transformant E. coli HB101 carrying pPLac 15 DNA were not different from that of L. casei as a donor strain on the basis of enzyme properites. However, specific activity of phospho-$\beta$-galactosidase in the cloned strain with Lac $Y^{-}$ phenotype of E. coli HB101 was lower than that in L. casei strain.

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