• Title/Summary/Keyword: Recombinant DNA

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Pseudomonas sp. W7로부터 Agarase 유전자의 Cloning 및 Escherichia coli에서의 발현

  • Ha, Jeong-Chul;Kim, Gu-Taek;Kim, Sung-Koo;Yu, Ju-Hyun;Kong, In-Soo
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.665-670
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    • 1995
  • A marine bacterium which produces extracelluar agarase was isolated from sea water. Isolated strain was identified as Pseudomonas sp. by the morphological and biochemical properties (1). HindIII restriction fragment of 3.2 kb from Pseudomonas genomic DNA was cloned into pUC19 to obtain recombinant plasmid pJA1 which enables E. coli JM83 to produce agarase. Most of agarase produced in E. coli was secreted into the culture medium. The enzyme (pJA1) showed the highest agarase activity during the stationary phase (20 hrs) of E. coli. The optimum temperature and pH were 40$\circ$C and 7.8, respectively. Restriction gene map anlaysis revealed that it has different restriction pattern with three kind of agarase gene reported.

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Genotoxicity Assay Using Chromosomally-Integrated Bacterial recA::Lux

  • Min, Ji-Ho;Gu, Man-Bock
    • Journal of Microbiology and Biotechnology
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    • v.13 no.1
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    • pp.99-103
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    • 2003
  • An Escherichia coli strain containing the recA promoter that fused to the luxCDABE operon originating from Photorhabdus luminescens was shown to respond sensitively to genotoxic stresses. Two different recombinant bacteria, one (DPDI 657) harboring a plasmid with the recA promoter that fused to the luxCDABE operon, and the other (DPD1710) containing a chromosomally-integrated recA promoter that fused with luxCDABE, were compared and it was found that the sensitivity of 'the two strains was significantly different in terms of their bioluminescent level, response time, and the minimum detectable concentration of a chemical causing DNA damaging stress. DPDI 710, with a chromosomally-integrated single copy, generally led to lower basal luminescence levels, faster responses, increased response ratios, and an enhanced sensitivity to mutagens, when compared to DPD 1657 with a multi-copy plasmid.

Lactobacillus casei YIT 9018의 Shuttle Vector 개발을 위한 분자유전학적 연구

  • Yoo, Min;Nam, Jin-Sik;Kwon, Oh-Sik;Baek, Young-Jin
    • Microbiology and Biotechnology Letters
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    • v.25 no.5
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    • pp.464-467
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    • 1997
  • A shuttle vector, pSHvec, was constructed for Lactobacillus casei (L. casei) YIT 9018 and JM1O9 by recombinant DNA technology. This vector contained the $\beta$-lactamase II gene from Bacillus cereus as a selection marker and replication origins for both Gram(+) and Gram(-) strains. It could transform the wild type L. casei YIT 9018 as well as E. coli JM109 and transformed cells were selected based on antibiotics resistance. The ability of L. casei YIT 9018 for curd formation in 11% skim milk was maintained even after transformation with pSHvec. The vector was stable as long as antibiotics were added to the medium. These results could contribute to the study of lactic acid bacteria for the industrial purpose at a genetic level.

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Cloning. Sequencing and Characterization of the Novel Penicillin G Acylase Gene from the Soil-isolated Leclercla adecarboxylata

  • Jun , Sang-O.;Lim, Ho-S.;Kim, Geun-Y.;Lee, Eung-S.;Lee, Mann-H.
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.331.3-332
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    • 2002
  • A novel penicillin G acylase (PGA)-producing bacterial strain was isolated from soil by using the Serratia marcescens overlay technique. The isolated strain was identified as Leclercia adecarboxylata based on the analyses of the biochemical characteristics (API 20E). the cellular fatty acid profile. and the 16S rDNA sequences. The gene encoding the PGA (pac gene) was cloned into the pHSG399 vector and the recombinant E. coliHB101 clones harboring the pac gene were isolated on agar plates containing phenylacetyl-L -leucine and penicillin G. (omitted)

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Parthenogenesis in Silkworm, Bombyx mori L.

  • Gangopadhyay Debnirmalya;Singh Ravindra;Kariappa B. K.;Dandin S. B.
    • International Journal of Industrial Entomology and Biomaterials
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    • v.10 no.1
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    • pp.1-10
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    • 2005
  • Parthenogenesis in mulberry silkworm, Bombyx mori L. acquires immense use in the development of outstanding homozygous lines with higher viability, hybrid vigour, combining ability and less phenotypic variability. It can serve as a powerful tool in controlling sex of the offsprings as well as a useful tool in selection. In fact India is the second largest silk producing country in the world next only to China and all the five types of natural silks viz., mulberry, oak tasar, tropical tasar, muga and eri are produced in India. However, little information is available on the role of artificial parthenogenesis in the development of superior silkworm breeds. This paper overviews some important studies carried out on artificial parthenogenesis, and outline of different types of parthenogenesis, methods of induction of artificial parthenogenesis, factors responsible for successful parthenogenetic development, cytogenetics of artificial parthenogenesis and role of artificial parthenogenesis in silkworm breeding. Besides, an attempt is made to describe briefly about parthenogenetic engineering which includes cloning in silkworm, artificial insemination, chimeras, hybridization, chromosomal substitution and recombinant DNA in silkworm.

Biotechnology for the Production of Threonine Production (Threonine의 생물공학적 생산)

  • Kim, Kyoung-Ja
    • YAKHAK HOEJI
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    • v.34 no.6
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    • pp.447-456
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    • 1990
  • Various methods are available for the production of L-threonine. The microbial production of L-threonine has been achieved by breeding L-threonine analog-resistant auxotrophic mutants of various bacteria. The enzymatic production of L-threonine has been demonstrated by use of threonine metabolic enzymes such as threonine deaminase, threonine aldolase, or threonine dehydrogenase complex. Threonine synthesis from glycine and ethanol seems to be catalyzed by the enzymes Methanol dehydrogenase(MDH) and Serine hydroxymethyltransferase(SHMT), which was also found to catalyze the aldol condensation of glycine with acetaldehyde. The improved production of L-threonine has been achieved by amplifying the genes for the L-threonine biosynthetic enzymes using recombinant DNA techniques.

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Current Status of the Research and Development of Bispecific Antibodies

  • Kwon, Sun-Il
    • Biomedical Science Letters
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    • v.26 no.3
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    • pp.136-148
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    • 2020
  • A bispecific antibody (BsAb) is an artificial protein containing two kinds of specific antigen binding sites. BsAb can connect target cells to functional cells or molecules, and thus stimulate a directed immune response. Last several decades a wide variety of bsAb formats and production technologies have been developed. BsAbs are constructed either chemically or biologically, exploiting techniques like cell fusion and recombinant DNA technologies. There are over 100 different formats of bsAb so far developed, but they could be classified into the two main categories such as Fc-based (with a Fc region) bsAbs and fragment-based (without a Fc region) bsAbs. BsAb has a broad application prospect in tumor immunotherapy and drug delivery. Here, we present a brief introduction to the structure of antibody, pharmacological mechanisms of antibodies and the trend in the production technologies of therapeutic antibodies. In addition, we address a review on the current status of various bsAb format development and their production technologies together with global situation in the clinical studies of bsAb.

A feruloyl esterase derived from a leachate metagenome library

  • Rashamuse, Konanani;Sanyika, Walter;Ronneburg, Tina;Brady, Dean
    • BMB Reports
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    • v.45 no.1
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    • pp.14-19
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    • 2012
  • A feruloyl esterase encoding gene (designated fae6), derived from a leachate metagenomic library, was cloned and the nucleotide sequence of the insert DNA determined. Translational analysis revealed that fae6 consists of a 515 amino acid poly-peptide, encoding a 55 kDa pre-protein. The Fae6 primary structure contained the G-E-S-A-G sequence, which corresponds well with a typical catalytic serine sequence motif (G-x-S-x-G). The fae6 gene was successfully over-expressed in E. coli and the recombinant protein was purified to 8.4 fold enrichment with 17% recovery. The $K_M$ data showed Fae6 has a high affinity to methyl sinapate while thermostability data indicated that fae6 was thermolabile with a half life ($T_{1/2}$) < 30 min at $50^{\circ}C$. High affinity for Fae6 against methyl sinapate, methyl ferulate and ethyl ferulate suggest that the enzyme can be useful in hydrolyzing ferulated polysaccharides in a biorefinery process.

New Concepts on Vaccine Development for the Poultry Diseases

  • Han, M.G.
    • Korean Journal of Poultry Science
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    • v.28 no.2
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    • pp.165-172
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    • 2001
  • Vaccination is one of the most important and cost-effective methods of preventing infectious diseases. Over the past decade, scientific in molecular biology and immunology have improved understanding of many diseases and led to the development of novel strategies for vaccination. An ideal vaccine would induce effective immunity specific for the type of infection, have long duration, require minimal or no boosters, have safety, would not induce adverse reaction, and be easy to administer. The desire to meet these criteria has resulted in the development of vaccines that do not depend on the use of the viable disease agent. It is not the intent of this review to give an extensive review of the field of vaccinology, but rather to address characteristics of conventional and genetically engineered vaccines.

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Production and Characterization of Monoclonal Antibodies to Yeast Mitochondrial RNA Polymerase Specificity Factor

  • Lee, Chang-Hwan;Jang, Sei-Heon
    • BMB Reports
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    • v.31 no.6
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    • pp.607-610
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    • 1998
  • Transcription of mitochondrial DNA in the yeast S. cerevisiae depends on recognition of a consensus nonanucleotide promoter sequence by mitochondrial RNA polymerase specificity factor, which is a 43 kDa polypeptide encoded by the nuclear MTF1 gene. Mtf1p has only limited amino acid sequence homology to bacterial sigma factors, but functions in many ways like sigma in that it is required for promoter recognition and initiation of transcription. To analyze the corebinding region of Mtf1p, monoclonal antibodies to this protein were prepared. Recombinant Mtf1p overproduced in E. coli was purified to near homogeneity and used to raise monoclonal antibodies (mAbs). From fused cells screened for Mtf1p mAbs by immunodot blot analysis, 19 positive clones were initially isolated. Further analysis of positive clones by Western blotting resulted in 4 mAbs of Mtf1p.

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