• Title/Summary/Keyword: 플라스미드

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Construction of Pretense-defective Mutant of Bacillus subtilis by Homologous DNA Recombination (상동성 유전자재조합을 이용한 단백질분해효소 비생산 바실러스균주의 구축)

  • Lee, Jin-Tae;An, Bong-Jeun
    • Food Science and Preservation
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    • v.7 no.4
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    • pp.414-417
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    • 2000
  • Competent cell transformation of B. subtilis AC819 was carried out using phenotypic protease-defective(Npr-) DNA of B. subtilis MT-2. An obtained transformant, designated B. subtilis HL-1, was obtained by homologous DNA recombination. Phenotypes of B. subtilis HL-1 were characterized histidine requirement streptomycin-resistance, tetracyclin resistance and non-producing protease. Protoplast transformation frequency of B. subtilis HL-1 by plasmid pUB110 was higher than that of B. subtilis MT-2. From this result, B. subtilis HL-1 is useful for protease gene transformation and thermostable protease gene cloning as a host.

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Studies on the Organization and Transcription of Aspergillus nidulans tRNA Genes (Aspergillus nidulans의 tRNA 유전자의 구성과 발현에 관한 연구 II. Aspergillus nidulans 총 tRNA 유전자의 cloning)

  • 이병재;강현삼
    • Korean Journal of Microbiology
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    • v.21 no.4
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    • pp.229-237
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    • 1983
  • Total tRNA genes from Aspergillus nidulans were cloned for the further investigation of the structure and expression of Aspergillus tRNA genes. Aspergillus DNA was isolated from spores and cloned into pBR322 plasmid DNA using BamHI and $T_4$ ligase. The recombinant hybrid DNA was transformed into E. coli HB101 and some 30,000 transformants were initially selected. Of these, about 5,300 E. coli clones containing Aspergillus DNA inserted into plasmid pBR322 at BamHl site have been isolated. The hybridization data obtained from the labeled Aspergillus $^{32}P-tRNA$ indicated that 105 colonies carried the total tRNA genes. From the data above and cohybridization experiment, tRNA genes of Aspergillus nidulans seem to be twice more clustered than those of yeast.

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Complete genome of a denitrifying Halioglobus sp. RR3-57 isolated from a seawater recirculating aquaculture system (순환여과양식시스템으로부터 분리된 Halioglobus sp. RR3-57의 유전체 분석)

  • Kim, Young-Sam;Noh, Eun Soo;Lee, Da-Eun;Kim, Kyoung-Ho
    • Korean Journal of Microbiology
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    • v.53 no.1
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    • pp.58-60
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    • 2017
  • Halioglobus sp. RR3-57 was isolated from a biofilter of a seawater recirculating aquaculture system and its complete genome sequence was obtained using the PacBio RS II platform. Two circular contigs were assembled and considered as a chromosome and a plasmid (size of 4,847,776 bp and 155,799 bp, and G+C content of 57.5% and 53.2%, respectively). Genomic analysis showed RR3-57 had 18 denitrification-related genes and an incomplete prophage.

Entrapment of Plasmid DNA in Liposomes (리포솜을 이용한 플라스미드 DNA의 봉입)

  • Song, Mi-Hyang;Lee, Mann-Hyung;Yong, Chul-Soon;Oh, Doo-Man
    • Journal of Pharmaceutical Investigation
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    • v.26 no.4
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    • pp.291-297
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    • 1996
  • Liposomes of $pSV-{\beta}-Galactosidase$ vector plasmid DNA with various lipid composition were prepared by the thin-film method. Size distribution, shape and the efficiency of plasmid DNA encapsulation were investigated. Effect of sonication time on the plasmid DNA entrapment in liposomes and stability at $4^{\circ}C$ were also examined. Sizes of neutral liposomes were about 100-200 nm and above $1\;{mu}m$, and those of cationic liposomes were about 400-600 nm and above $1\;{mu}m$. Shapes of liposomes entrapped plasmid DNA were spherical. Proper sonication time for better entrapment was below 15 minutes and stability at $4^{\circ}C$ was decreased rapidly after 1 day. Plasmid DNA entrapments of complex liposomes of various lipids were higher than those of liposomes made from one sort of lipid. Plasmid DNA entrapments of cationic liposomes were higher than those of neutral liposomes.

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Expression of Fusion Products of Insecticidal Crystal Protein Genes from Two Different Bacillus thuringiensis Strains (두종의 Bacillus thuringiensis 내독소단백질 유전자의 융합에 의한 발현)

  • 제연호;김상현
    • Journal of Sericultural and Entomological Science
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    • v.35 no.1
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    • pp.36-42
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    • 1993
  • Expression of insecticidal protein by fusion product of truncated HD-1[CryIA(a)] N-terminal and HD-73[CryIA(c)] C-Terminal fragment of Bacillus thruingiensis subsp. kurstaki was investigate. Immunological analysis of transformants by using polyclonal antisera raised against the whole-crystal protein of HD-1 revealed that SK4 and SK5 were observed cross-reaction with polypeptides of 77-kDa and 105-kDa, respectively. Bioassay of the transformant pSK5 to Plutella maculipennis and Heliothis assulta were 96% and 97%, respectively.

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Generation and Expression of Amino-Terminal Domain of the Gene Coding for the Lumazine Protein from Photobacterium phosphoreum (발광 박테리아 Photobacterium phosphoreum의 Lumazine Protein을 코드 하는 유전자의 염기 서열 분석 및 발현)

  • Woo Young-Eun;Kim So-Young;Lee Chan-Yong
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.306-311
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    • 2005
  • In this study, the amino-terminal half truncated lump and the whole lump genes from Photobacterium phosphoreum coding for the lumazine protein were cloned by polymerase chain reaction and expressed in Escherichia coli. To identifiy of the binding site of the ligand or substrate, the amino acid identities from the sequences of the lumazine protein, yellow fluorescent protein, and riboflavin synthase from different organisms were also compared and analyzed.

Complete genome sequence of Pseudoalteromonas donghaensis HJ51T isolated from seawater (해수에서 분리된 Pseudoalteromonas donghaensis HJ51T 의 유전체 서열분석)

  • Oh, Ji-Sung;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.54 no.3
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    • pp.305-307
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    • 2018
  • The whole genome sequencing using PacBio RS II platform was performed for a marine bacterium Pseudoalteromonas donghaensis $HJ51^T$ isolated from East Sea of Korea. As a result, three assembled contigs consisting of a chromosome (size of 3,646,857 bp, and G + C content of 41.8%) and two plasmids (size of 842,855 bp and 244,204 bp, and G + C content of 41.3% and 40.4%, respectively) were obtained. The genome included 4,083 protein coding genes and 127 RNA genes. This result could be used for gene sources of biopolymers degradation and the development as a new host with secretion system similar to Escherichia coli.

Development of Bactericidal Yeast Strain by Expressing the Leucocin A Gene on the Cell Surface of Saccharomyces cerevisiae (Saccharomyces cerevisiae세포 표면에 leucocin A유전자의 발현에 의한 항균활성 효모의 개발)

  • Lee Sang-Hyeon
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.923-927
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    • 2005
  • ln order to develop yeast cells that produce a bacteriocin on their cell surfaces, the 114 bp Leucocin A gene with stop codon was ligated into pYDl, an yeast vector. The recombinant DNA, pYDl-LeucoA was used to transform yeast (Saccharomyces cerevisiae) cells. Yeast cells harboring pYDl-LeucoA showed antibacterial activity against Bacillus subtilis. To confirm these bacteriocidal yeast cells possess the Leucocin A gone, PCR was performed with plasmid prepared from transformed yeast cells as a template and two Leucocin A-specific primers. In this study, bacteriocidal yeast cells that can be used as an antibiotic or a food preservative were developed.

Studies of Coagulase Production and Isolation of R-plasmid from Staphylococcus aureus (Staphylococcus aureus의 Coagulase 생성능과 R-플라스미드 분리에 관한 연구)

  • Yoon, Hyo-Sook;Lee, Hyung-Hoan;Kim, Soo-Young
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.3
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    • pp.259-266
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    • 1987
  • A total of 129 clinical isolates of Staphylococcus species was characterized by the tests of coagulase production, haemagglutination, mannitol fermentation, DNase production and hemolysis. Ninety-nine out of them showed positive reactions to the tests, therefore they were identified as Staphylococcus aureus. The isolates showing positive reaction in haemagglutination test also showed 100% of tube coagulase positive reaction. The haemagglutination test was a reliable method for identifying Staphylococcus aureus in the clinical laboratory. S. aureus produced stronger hemolysis with human blood agar than with sheep blood agar. Antibiotic resistant S. aureus isolates(S-46, S-112, S-126) had 4 to 6 p]asmid DNA elements. The S-112 strain had 6 plasmid DNA elements(1.8, 2.2, 3.7, $26.3{\sim}50$, and 70 Mdaltons), the S-126 had 4 elements(2.6, 4.2, $4.6{\sim}60Md$), and the S-46 had 1 element(${\sim}100Md$). PPSA strain had 4 plasmid DNA elements(2.5, 4.2, $4.6{\sim}60Md$) and S. aureurs(ATCC) strain contained 9.4, 26.3 and ${\sim}50Md$ plasmid DNA elements.

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Complete genome sequence of Bacillus aryabhattai K13 isolated from compost (퇴비에서 분리한 Bacillus aryabhattai K13의 유전체 염기서열)

  • Patil, Kishor Sureshbhai;Lee, Seung-Je;Le, Ve Van;Jeon, So Hyun;Chae, Jong-Chan
    • Korean Journal of Microbiology
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    • v.53 no.4
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    • pp.332-333
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    • 2017
  • Bacillus aryabhattai K13 was originated from enriched culture of compost using 1% kraft lignin as a sole carbon and energy source. The complete genome of strain K13 consists of one 5.0 Mb chromosome and two circular plasmids with 139 kb and 78 kb, respectively. Analysis of the genome determined in this study may contribute to identify the genes responsible for degradation of lignin.