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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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The Korean Journal of Microbiology
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Journal DOI :
The Microbiological Society of Korea
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Volume & Issues
Volume 28, Issue 4 - Dec 1990
Volume 28, Issue 3 - Sep 1990
Volume 28, Issue 2 - Jun 1990
Volume 28, Issue 1 - Mar 1990
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Molecular Cloning and High-Level Expression of Human Cytoplasmic Superoxide Dismutase Gene in Escherichia coli
The Korean Journal of Microbiology, volume 28, issue 2, 1990, Pages 91~97
Complementary DNA (cDNA) coding for human cytoplasmic superoxide dismutase (SOD1) (superoxide: superoxide oxidoreductase E.C.184.108.40.206) was isolated from human liver cDNA library of
gt11 by in situ plaque hybridization. The insery cDNA gas the 5' untranslational region (UTR) and 3'UTR of SOD1 gene. Polymerase Chain Reaction (PCR) method was used fro subcloning of SOD1 structural gene. Using synthetic sense strand primer (24mer) containing a start codon and antisense strand primer (24mer), SOD1 structural gene was selectively amplified. Amplified DNA was directly cloned into the HincII site of pUC19 plasmid. Insery cDNA was subcloned into M13 mp19 and sequenced by dideowy chain termination method with Sequenase. The nucleotide sequence of insert cDNA had an open reading frame (ORF) coding for 153 amino acid residues. The structural gene of cytoplasmic SOD was placed under the control of bacteriophage
regulatory sequences, generating a highly efficient expression plasmid. The production of human SOD1 in E. coli cells was about 7% of total cellular proteins and recombinant human SOD1 possessed its own enzymatic acitivity.
Genetic Recombination of Brevibacterium lactofermentum by Protoplast Fusion
The Korean Journal of Microbiology, volume 28, issue 2, 1990, Pages 98~103
Brevibacterium lactofermentum SWA (arg trp) and B. lactofermentum SWB (met ser) were obtained from UV and NTG treatment. The rates of protoplast formation by B. lactofermentum SWA and SWB were 99.93% and 99.98%, respectively when each strain was treated with penicillin G in mid exponential growth phase, followed by incubation with 400
/ml of lysozyme in lysis fluid supplemented with 0.4M sucrose. Frequencies of protoplast regeneration in B. lactofermentum SWA and B. lactofermentum SWB were 9.27% and 10.32% respectively, on regeneration medium containing 0.5M sodium succinate, 50 mM
, and 3% PVP. In intraspecific protoplast fusion between B. lactofermentum SWA and B. lactofermentum SWB, fusion frequency of
was observed by using the 100mM
and 30% PEG 6,000 in fusion fluid. Relative recombinant frequencies in each marker by means of selective media could be used for genetic analysis.
Cloning of agrobacterium tumefaciens chromosomal virulence region
;Cangelosi, G.A.;Nester, E.W.;
The Korean Journal of Microbiology, volume 28, issue 2, 1990, Pages 104~108
The chromosomal DNA of Agrobacterium tumefaciens contains the genes required for bacterial attachment to plant cell which is an essential atage in crown gall tumorigenesis by Ti-plasmid. In order to clone the genes, Agrobacterium tumefaciens strain A5512 was mutagenized by transposon Tn5 and two Agrobacterium tumefaciens mutants which are attachment-defective and nontumorigenic were isolated. From one of the two mutants, a chromosomal virulence region which was required for attachment to the plant cells was cloned.
Mutator effects of plasmid pKM101 and pSL4 to E. coli DNA repair
The Korean Journal of Microbiology, volume 28, issue 2, 1990, Pages 109~113
The mutagenesis-enhancing plasmid pKM101 and its mutant pSL4 were introduced into Escherichia coli B/r strains possessing different DNA repair capacities (
) and determined the protection effect and mutagenecity for UV and MNNG. The mutability and protection effect of plasmid pKM101 and pSL4 were affected by different DNA repair capacity. The mutagenecity and resistance of two plasmids were increased against UV and MNNG, and plasmid pSL4 had a higher effect than pKM101. We suggest that the functional differences between pKM101 and pSL4 is due to the variety of mutator gene.
Expression of a Yeast Superkiller Gene(SK13) in Saccharomyces cerevisiae
;Wickner, Reed B.;
The Korean Journal of Microbiology, volume 28, issue 2, 1990, Pages 114~119
A yeast chromosomal superkiller gene (SK13) was cloned and expressed in
Saccharomyces cerevisiae strains. The gene was fused to the structural region of E. coli lacZ gene at its C-terminus in a yeast-E. coli shuttle vector, pSR605. The fused gene complemented
strains with SK13 activity and the quantitative level of expression was measured as determined by assaying
-galactosidase activity. The SDS-polyacrylamide gel electrophoresis and the Western blot analysis of this fused protein showed the immuno-reacted bands with a protein of the estimated molecular size (ca.250Kd).
Construction of Starch-Fermenting Yeast Using Protoplast Fusion Technique
The Korean Journal of Microbiology, volume 28, issue 2, 1990, Pages 120~127
Intraspecific fusion frequency between Filobasidium capsuligenum CBS6122-2 ade and met trp or met strains was
, respectively. And intergeneric fusion frequency between Folobadidium capsuligenum CBS 4318 (cys his) and Saccharomyces cerevisiae 262-12-1 (hom3 thr1) or X2180-1A (ade thr) was
, respectively. Nuclear fusion appears to occru in fusants between intraspecies and intergenera as strongly suggested by DNA content, nuclear staining, comparison of survival rate to UV light and mitotic segregation analysis. It was also found that
-amylase and glucoamylase activity from intraspecific hybrids was 1.6-2.1 fold increased when compared with that from thier parents.
Studies on the Histones of the Genus Rhizopus
The Korean Journal of Microbiology, volume 28, issue 2, 1990, Pages 128~133
The chromatin of all higher eukaryotic cells contains a group of very basic low-mole-cular weight proteins, the histones. But much less is known about histones in lower eukaryotes. Our purpose was to study the histones of the genus Rhizopus. After isolation and purification of nucleoprotein the basic nucleoproteins were analyzed by gel electrophoresis, in sodium dodecyl sulfate as well as acid/urea gels and compared with calf thymus histones. Their electrophoretic mobility in polyacrylamide gel indicate that they are histone homologous, although not identical, to the H2A, H2B, H3 and H4 histones of mammals with the exception of H1. The result suggests that Rhizopus thus appears to contain histone proteins which are homologous to the histones from in higher eukaryotes. The similarity between the calf thymus histone H1 and the Rhizopus high band group remains to be discussed.
Cadmium Detoxification Mechanism in Klebsiella aerogenes ATCC 10031
The Korean Journal of Microbiology, volume 28, issue 2, 1990, Pages 134~144
In order to examine that what kind of system correlated with cadmium detoxification mechanism in Klebsiella aerogenes ATCC 10031, we tried to investigate the effect of phosphate upon the detoxification and also elucidate whether the cadmium phosphate and/or polymeric Cd-Pi complex is formed actually in cell or not. As the results, it was shown that growing pattern had long lag adaptive phase of 12 hr to 24 hr, at the concentrations of 0.02 mM and 0.08 mM cadmium, respectively. Cadmium was accumulated more highly in the fraction of cell wall and membrane than in those of cytoplasm. In case of phosphate starving cells added cadmium, inorganic polyphosphate system was primarily correlated with Cd-detoxification during the lag phase for the accommodation to cadmium, on the other hand, Cd:Sulfide complex system secondarily correlated it during the stationary phase. These results implied that polyphosphate system and Cd:sulfide complex system, these two systems were operated compensatively each other. Considering the results obsdrved with EM and examined tha changes of sulfide and polyphosphate amount, it was reflected that Cd:S complex was located at the cell surface. In the results of
P NMR spectra in the cells with cadmium pressure, several phosphate signals arose newly from the polyphosphate region with moving chemical shift of it. This phinomenon strongly implied the actual existence of Dd:Pi comples and /or Cd:poly-P complex in the cell and also the cellular compartmentalization of cadmium detoxifying mechanism.
Effects of Environmental Factors on Degradation of Aroclors by Gram-negative Bacteria
The Korean Journal of Microbiology, volume 28, issue 2, 1990, Pages 145~150
The effects of environmental factors on degradation of Aroclor 1242 were investigated with four Gram-negative bacterial isolates. Their biodegradabilities of the Aroclor were well correlated to their growth rates on the Aroclor added as a sole carbon and energy source. The optimum concentration of the Aroclor for biodegradation of the substrate in MM2 medium was 0.5mg/ml in HK-100, HK-123, and MS-1003 strains, but 1 mg/ml in DJ-26 strain. The optimum temperature and pH were
and 7.0, respectively, for all the strains. On the basis of the results which the strain of DJ-26 showed the highest degradability of the Aroclor as well as the highest growth rate under the optimum environmental conditions, the bacterial isolate identified as Pseudomonas sp. was found to be a strain usable for treatment of the toxic and recalcitrant chemical pollutants, such as polychlorinated aromatic hydrocarbons.
Properties and Activities of Nireogenase System of Azospirillum amazonensa Kp1
The Korean Journal of Microbiology, volume 28, issue 2, 1990, Pages 151~157
The maximum nitrogen fixation activity of the associative, microaerobic and acid tolerant bacteria, Azospirillum amazonense Kp1 was obtained with 0.2Kpa of
and showed a reversible inhibition by the higher concentrations. Ammonium treatment caused a gradual inhibition of the activity up to 350mM. The nitrogenase systems were purified by gradient chromatography on DEAE-52 cellulose, heat treatment and preparative PAGE. The MoFe protein showed molecular weight of 210,000 including two nonidentical subunits with apparent molecular weights of 55,000 and 50,000 and an isoelectricpoint of 5.2 and contained 2, 24 and 28 atoms of Mo, Fe and acid labile S per molecule. The Fe protein revealed molecular weight of 66,000 including two types of subunits with molecular weights of 35,000 and 31,000 and an isoelectric point of 4.6, and contained 4 atoms of Fe and 6 atoms of S per molecule. The maximum specific nitrogenase activity attained 2,200 and 1,700nM
, respectively for MoFe and Fe proteins at pH7 and
. The activity was lost after 10 and 30 days under the cold room (
) condition for Fe and MoFe proteins, respectively.
An Improved Method for the Measurement of Fungal Degradability of Synthetic Polymers
The Korean Journal of Microbiology, volume 28, issue 2, 1990, Pages 158~161
An improved fungal test method, soft agar overlay method, has been developed for the measurement of biodegradability of synthetic polymers. Advantages of this technique over conventionally used test method of ASTM include better visualization of fungal growth on polymeric materials, shortening of incubaiton period, and engnaced sensitivity especially to polymeric materials containing highly recalcitrant molecules.
Effects of Culture Environments on Alkaline Protease Biosynthesis in Streptomyces sp.
The Korean Journal of Microbiology, volume 28, issue 2, 1990, Pages 162~168
The aims of the present study were to evaluate the effects of culture conditions on the biosynthesis of extra-cellular alkaline protease in Streptomyces sp. The formation of aerial mycelia and spores were compared with the protease production in order to know the relations between the alkaline protease and the cell differentiation. As results, it was found that substrate concentration was very critical to regulate the formation of the protease, aerial mycelia, and spores, which were resulted from the changes of culture pH to acid. When the culture pH was adjusted with phosphate buffer from pH 6 to pH 9, the alkaline protease production was increased as the culture pH increased whereas aerial mycelia and spore formation were reversely related to the culture pH. Therefore, it was thought that the culture pH was an important factor to regulate the alkaline protease synthesis.
Construction of Plasmids for Overproduction of L-Phenylalanine
The Korean Journal of Microbiology, volume 28, issue 2, 1990, Pages 169~173
For the overproduction of L-phenylalanine using Escherichia coli, the authors constructed various recombinant plasmids including pMW 10, pMW 11 and pMW 12. The
genes for the production of L-phenylalanine were isolated from Escherichia coli MWEC 101-5 strains. The productivity and atability of Escherichia coli regulatory mutants containing recombinant plasmids were investigated to evaluate the efficiency of the
genes. The MWEC 101-5/pMW 11 strain produced 24.3g/l of L-phenylalanine while its stability was 73.8 percent. The specific activity of prephenate dehydratase in the MWEC 101-5/pMW 11 strain increased by 26-fold compared with that of Escherichia coli K-12.
L-Phenylalanine Production by Regulatory Mutants of Excherichia coli K-12
The Korean Journal of Microbiology, volume 28, issue 2, 1990, Pages 174~179
In order to overproduce L-phenylalanine, various kind of regulatory mutants were isolated from parental Escherichia coli K-12. MWEC 83 Producing 7.4g/l of L-phenylalanine has been derived as a tyrosine and tryptophan double auxotrophic mutant. To produce L-phenylalanine without adding L-tyrosine and L-tryptophan, revertant strain MWEC 101 was isolated from MWEC 83. Further various analogues and valine resistant mutants were isolated from MWEC 101. MWEC 101-5 was the most excellent strain that produced 17.9g/l of L-phenylalanine after having been cultivated for 54 hours in 15% glucose medium. It was disclosed that activities of rate-limiting enzymes including chorismate mutase and prephenate dehydratase in MWEC 101-5 were desensitized to 2mM L-phenylalanine in the enzyme reaction mixture and that activities level of 3-deoxy-D-arabino-heptulosonic acid-7-phosphate synthase and prephenate dehydratase were increased more than 20 times over those of the parental strain.