• Title/Summary/Keyword: Plasmid DNA

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Replication origin (ori) of R-plasmid pSBK203 Isolated from Staphylococcus aureus DHI (Staphylococcus aureus DH1에서 분리한 R-plasmid pSBK203상의 복제개시 부위 ori에 관한 연구)

  • Min, Kyung-Il;Byeon, Woo-Hyeon
    • Korean Journal of Microbiology
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    • v.32 no.3
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    • pp.186-191
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    • 1994
  • The origin of the leading strand replication (ori) and of lagging strand replication (M-O) of R-plasmid pSBK203 was identified and its base sequence was determined. About 50 bp of ori sequence residues overlapped with the structural gene of rep. Sequence comparison reveals that pSBK-ori shares obvious identities with those of pT181 family and consists of two regions, one is conserved and the other is variable region. Of two palindrome sequence located one after another in upstream region of rep gene, palA' instead of palA which shares sequence homology with diverse family of plasmids such as pOX6, pC194, and pE194 seems to act as a signal for conversion of primarily replicated ssDNA to dsDNA (minus origin (M-O)).

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Studies on the Effective Drug Delivery System Using Naked Plasmid DNA for the Erythropoietin Expression in vivo

  • Park, Yeong-Seop;Jeong, Dong-Geon;An, Jin-Ho;Choe, Cha-Yong;Ju, Hyeon
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.582-586
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    • 2003
  • There has been interest in developing gene therapy based on naked plasmid DNA for treating serum protein deficiencies and human erythropoietin (hEPO) is one of the candidate for gene therapy being Investigated most enthusiastically. We constructed novel plasmid DNA vectors pVAC-hEPOI/II/III which contain one, two, three hEPO gene(s) respectively for producing high level expression and secretion of hEPO in vitro and in vivo. NIH3T3 and COS7 cells were transfected transiently with these vectors and increase in hEPO expression in medium reached 2-5 fold in comparison with pSecTagB-hEPO. Intra muscular administrations of pVAC-hEPOI/II/III vectors into mice resulted in high level secretion of hEPO in the serum and corresponding increases in hematocrit level. In conclusion the transduction efficiency of naked plasmid vectors is one of the critical factors of a gene delivery system and these novel plasmid vectors will contribute to various gene therapy based on naked plasmid DNA.

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Molecular Cloning and Expression of Bacillus pasteurii Urease Gene in Escherichia coli (B. pasteurii Urease 유전인자의 E. coli의 복제와 발현)

  • Kim, Sang-Dal;John Spizizen
    • Microbiology and Biotechnology Letters
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    • v.13 no.3
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    • pp.297-302
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    • 1985
  • The 7.1 Mdal Xbaf fragment of Bacillus pasteurii ATCC 11859 containing gene for urease was inserted into the Xbal site of bifunctional plasmid pGR71, and its urease gene was cloned and expressed in E. coil RRI. But the cloned gene was not expressed in Bacillus subtilis BR151 in consequence of deletion of inserted DNA fragment. The recombinant plasmid thus formed was named pGU66. The restriction map of the plasmid pGU66 was determined, and the size of the plasmid was estimated to be 12.6 Mdal by double digestion of restriction enzymes of the plasmid. The urease of the cloned strain was accumulated in periplasmic space and very similiar to that of donor strains in their enzymatic properties.

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In Vivo Kinetics and Biodistribution of a HIV-1 DNA Vaccine after Administration in Mice

  • Kim, Byong-Moon;Lee, Dong-Sop;Choi, Jae-Hoon;Kim, Chae-Young;Son, Mi-Won;Suh, You-Suk;Baek, Kwan-Hyuck;Park, Ki-Seok;Sung, Young-Chul;Kim, Won-Bae
    • Archives of Pharmacal Research
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    • v.26 no.6
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    • pp.493-498
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    • 2003
  • In this study we have investigated the pharmacokinetics and tissue distribution of GX-12, a multiple plasmid DNA vaccine for the treatment of HIV-1 infection. Plasmid DNA was rapidly degraded in blood with a half-life of 1.34 min and was no longer detectable at 90 min after intravenous injection in mice. After intramuscular injection, plasmid DNA concentration in the injection site rapidly declined to less than 1 % of the initial concentration by 90 min post-injection. However, sub-picogram levels (per mg tissue) were occasionally detected for several days after injection. The relative proportions of the individual plasm ids of GX-12 remained relatively constant at the injection site until 90 min post-injection. The concentration of plasmid DNA in tissues other than the injection site peaked at 90 min post-injection and decreased to undetectable levels at 8 h post-injection. The rapid in vivo degradation of GX-12 and absence of persistence in non-target tissues suggest that the risk of potential gene-related toxicities by GX-12 administration, such as expression in non-target tissues, insertional mutagenesis and germline transmission, is minimal.

Cloning of Bacillus amyloliquefaciens amylase gene using YEp 13 as a vector II. Expression of cloned amylase gene in Saccharomyces cerevisiae (YEp 13 vector를 이용한 Bacillus amyloliquefaciens amylase gene의 cloning II. Saccharomyces cerevisiae에서의 발현)

  • 김관필;서정훈
    • Microbiology and Biotechnology Letters
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    • v.14 no.3
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    • pp.209-212
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    • 1986
  • $\alpha$-Amylase gene of Bacillus amyloliquetaciens was cloned on plasmid YEp13, S. cerevisiae-E. coli shuttle vector. Hybrid plasmid pTG17, carrying $\alpha$-amylase gene of B. amyloliquefaciens, was transformed to E. coli and the expression of it in yeast was investigated. This plasmid was unstable in E. coli and produced two minor plasmids, pTG17-1 and PTG17-2, which resulted from the segregation of it. Transformant of S. cerevisiae MC16 with pTG17-1 plasmid was not appeared on SD medium because of the Leu2 gene defection. S. cerevisiae could be transformed by the hybrid plasmid, and $\alpha$-amylase activity of the yeast transformant was detected by somogyi-Nelson method and agar diffusion method.

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Application of in situ gelling mucoadhesive delivery system for plasmid DNA as a macromolecule

  • Park, Jeong-Sook;Oh, Yu-Kyoung;Kim, Chong-Kook
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.236.1-236.1
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    • 2002
  • Mucosal administration of drug or therapeutic gene is emerging as a new route of delivery for systemic and local therapeutics. Previously. in situ gelling system has been applied to chemical drug such as acetaminophen. insulin. prostaglandin E1. and clotrimazole. Plasmid DNA has not been delivered in form of in situ gelling vehicles. To improve the intranasal absorption of plasmid DNA. we designed delivery systems composed of provide of in 냐셔 gelling and mucoadhesive polymers. (omitted)

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Drug Resistance in Fish-Pathogenic Bacteria

  • Aoki, Takashi
    • Journal of fish pathology
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    • v.6 no.1
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    • pp.57-64
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    • 1993
  • The properties and DNA structures of R plasmids differ depending on the species of the fish-pathogens Aeromonas hydrophila, A. salmonicida, Edwardsiella tarda, Enterococcus seriolicida, Pasteurella piscicida and Vibrio anguillarum. However, some R plasmids with the same resistance markers in similar DNA structures were found in A. hydrophila and E. tarda, as well as in A. hydrophila and A. salmonicida. R plasmids from V. anguillarum were classified into three groups according to their DNA structures. The first group was detected before 1977, the second from 1980 to 1983, and the third from 1989 to 1991. R plasmids have been retained within P. piscieida having the same DNA structures and detected at various locations and times. E. seriolicida strains carrying the same R plasmids, which were encoded with resistance to macrolide antibiotics(MLs), lincomycin(LIM), and TC, and to MLs, LIM, and CP. were distributed in yellowtail farms in various districts. The chloramphenicol-resistance(cat) gene of the R plasmids of P. piscicida was classified as CAT type I. The cat of the R plasmids of E, tarda. A. salmonicida was classified as type II. The cat of R plasmids of V. anguillarum was classified into two types. One type detected before 1977, was classified as CAT IV and the other type, detected after 1980, was classified as CAT II. Tetracycline-resistance (tet) V. anguillarum, isolated before 1977 and after 1981, was classified as Tet B and Tet G, respectively. The class D tet gene was widely distributed in R plasmids from fish-pathogens A. hydrophila, E. tarda, P. piscicida, and also V. anguillarum isolated after 1989.

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A Small Cryptic Plasmid pZMO1 of Zymomonas mobilis ATCC10988

  • Kang, Hyung-Lyun;Kang, Hyen-Sam
    • Genomics & Informatics
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    • v.1 no.1
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    • pp.55-60
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    • 2003
  • The nucleotide sequence of pZMO1, a small cryptic plasmid of Zymomonas mobilis ATCC10988 was determined. Analysis of 1,680 bp of sequence revealed $69\%$ identity with Shigella sonnei plasmid, pKYM and $61\%$ identity with Nostoc sp. ss DNA replicating plasmid. Analysis of a deduced amino acid sequence of an orf of pZMO1 revealed $75\%$ identity and $90\%$ similarity with the repA gene of Synechocystis sp. plasmid pCA2.4. The upstream region of the repA gene of pZMO1 possesses six directed repeat sequences and two inverted repeat sequences at downstream of the IR consensus sequence of nick region of rolling circle replication (RCR) plasmid. A typical terminator hairpin structure was found at the downstream region of repA gene. Degradation of single-stranded plasmid DNA by S1 nuclease was detected by Southern hybridization. It suggests that pZMO1 replicates by a rolling circle mechanism in Z. mobilis ATCC10988 cells.

Molecular Cloning of a CMCase Gene from Alkalophilic sp. and Its Expression in Escherichia coli

  • Yu, Ju-Hyun;Kong, In-Soo;Kim, Jin-Man;Park, Yoon-Suk
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 1986.12a
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    • pp.529.1-529
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    • 1986
  • For isolation of the CMCase gene of the alkalophilic Bacillus sp. strain N-4 to analyze their genetic information for the multicomponents of the cellulase, Bscherichia coli K12 and plasmid DNA pBR322 was used as host-vector system. After the digestion of purified chromosomal DNA and plasmid DNA pBR322 with HindIII, these were ligated. The ligated DND were transformed into Escherichia coli, and recombinant plasmid 107 carried the gene coding for CMCase was constructed. The CMCase produced by Escherichia coli cells containing plasmid DNA pYBC107 was found in the cells as intracellular enzyme and nearly 60% of the total CMCase activity was localized in cellular fraction. Also, the optimum pH for the reaction of CMCase produced by Escherichia coli was appeared at pH .8.0 and the enzyme was stable between pH 7.0 and pH 8.0.

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Isolation of Plasmid DNA in Pediococci from Kimchi (김치의 Pediococci에 존재하는 Plasmid DNA 분리)

  • Park, Yun-Hee;Ryu, Uk-Sang;Jo, Do-Hyun
    • Applied Biological Chemistry
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    • v.31 no.1
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    • pp.33-37
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    • 1988
  • Three species of Pediococci, Pediococcus pentosaceus, Pediococcus acidilactici and Pediococcus halophilus were isolated from Kimchi. P. pentosaceus and P. acidilactici showed inhibitory activity against Streptococcus faecalis, Pseudomonas sp., P20 and Vibrio parahaemolyticus. However, the growth of all test organisms was not inhibited by P. halophilus. Ten strains contained one to seven plasmids, ranging in size from 1 to 60 megadaltons.

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