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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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The Korean Journal of Microbiology
Journal Basic Information
Journal DOI :
The Microbiological Society of Korea
Editor in Chief :
Volume & Issues
Volume 27, Issue 4 - Dec 1989
Volume 27, Issue 3 - Sep 1989
Volume 27, Issue 2 - Jun 1989
Volume 27, Issue 1 - Mar 1989
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Expression and Secretion of Hepatitis B Viral Mutant Core Antigen
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 169~175
To study the role of mutant precore region in expression and secretion of hapatitis B viral core antigen, we have cloned core antigen gene(HBc) with or without precore region in geterologous expression vectors containing SV40 promoter, yeast promoter, and lambda
promoter. In COS cells transfected with plasmid containing C-gene with precore region, antigens were detected in both cell extract and cultured medium. However, in the cells transfected with plasmids containing C-gene without precore or with mutated precore region by one nucleotide (T) addition at the nucleotide 1,821, HBcAg was detected only in cell extracts. These results support that the mutation by one nucleotide addition shifted the initiation codon of precore region to 53 nucleotides upward and the elongated precore region also played a major role in the secretion of HBcAg in mammalian cells. In the case of yeast and E. coli, HBcAg was detected only in cell extracts in spite of the presence of precore region, which suggest that precore region could not affect HBcAg secretion in these system.
Site-Specific Mutagenesis on the 32-T and 39-T of E. coli
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 176~180
There are three pseudourdine (
)bases in the E. coli
In order to study the function of the pseudouridine bases in the
, changes of bases
gene to other bases were undertaken by the site-specific mutagenesis. Site-specific mutagenesis of T in the pheW gene, a
gene of E. coli, corresponding to the baseat the No.32 position to C and also T corresponding to the base at the No.39 position to C were performed using Kunkel's uracil-containing template method. Identification of mutants were undertaken by the KNA sequencing techniques of the mutated pheW genes and activities of the mutated pheW genes complementing to E. coli NP37 mutant(
) using the recombinant plasmid containing the mutated genes. Neither NP37 harboring pheW gene mutated at No.32 position nor NP37 harboring pheW gene mutated at No.39 position can be grown at non-permissive temperature. The result means that both mutated pheW genes can not complement to E. coli NP37, and that the pseudouridine bases are essential to the activity of the E. coli
-galactosidase gene located on plasmid in lactobacillus casei
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 181~187
Plasmid DNA was isolated from Lactobacillus casei SW-M1(
strain). The curing frequencies of pPLac plasmid from L. casei SW-M1 showed 43% for acriflavin treatment and 53% for ethidium bromide treatment after 3 times transfer. On the charaterization of pPLac plasmid, it was found that the plasmid contained gene encoding phospho-
-galactosidase for lactose utilization. Lactose-PTS(phosphotransferase system)was involved in membrane transport system in
strain. Induction of phospho-
-galactosidase was specially effective by galactose, lower effect with lactose and glucose but not by IPTG(isopropyl-
-D-thiogalactoside). This result showed that induction of phospho-
-galactosidase by IPTG did not appeared. The catabolite repression of phospho-
-galactosidase synthesis by glucose was not found in L. casei.
Molecular cloning of phospho-
-galactosidase gene of lactobacillus casei in escherichia coli
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 188~193
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-
-galactosidase by the determination of enzyme activity. Phospho-
-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-
-galactosidase in the cloned strain with Lac
phenotype of E. coli HB101 was lower than that in L. casei strain.
Nucleotide Sequence and Inducibility Analysis of Chloramphenicol Acetyltransferase Gene from Staphylococcus aureus R-plasmid pSBK203
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 194~200
The nucleotide sequence of inducible chloramphenicol acetyl-transferase(CAT) gene isolated from a small plasmid pSBK203 of Staphylococcus aureus was determined. The base sequence shows that structural gene of pSBK203-CAT encodes a protein of 213 amino acids and has a leader region which encodes a short polypeptide of 9 amino-acids in its upstream. vertical bar /sup 35/S vertical bar-Methionine labelled CAT gene product in minicell showed almost same mobility with pC194-CAT of which molecular weight is 24Kdal on polyacrylamide gel electrophoresis. Predicted amino acid sequence of pSBK203-CAT has revealed a high degree of homology with the CATs of pC194 and pC221 than those of cat-86, Tn9 and proteus mirabilis PM13.
A study of aerobic and anaerobic inducible genes using Mu dJ(Km lac) operon fusion in salmonella typhimurium
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 201~209
Using Mu dJ(Km lac) operon fusion, several oxygen-regulated genetic loci were identified in Salmonella typhimurium. Nine aerobically inducible(oxi) and thirteen anaerobically inducible(ani) operon fusions were newly identified. Based on the control by oxrA regulatory locus, the ani-lacZ fusions were grouped into two classes: class I loci were regulated by oxrA regulatory locus; class II genes were not affected by this locus. Some of the ani-lacZ fusions had required growth in CAA and LB before they exhibited the inducible phinotype. Most of all ani-lacZ fusions were repressed by nitrate and fumarate. Three of the ani loci were mapped into
map unit (YK114),
map unit(YK120), and
map unit(YK112) by testing the cotransduction frequency, respectively.
Genes involved in mating processes of saccharomyces cerevisiae
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 210~215
In order to elucidate and characterize the signal transduction pathway(s) whereby yeast cells respond to mating pheromone, we have isolated mutants which are able to conjugate in the absence of the alpha-factor receptor. Sixty-one suppressors of a ste2-deletion mutation which also confer a ts conditional "start" arrest phenotypw have been subjected to genetic analysis. The mutants could be assigned to three complementation groups designated CDC70, CDC72 and CDC73, which are unlinked to each other as well as to the previously identified start genes. Quantitation of mating ability of the cdc70, cdc72 and cdc73 mutations in a ste2-deletion background gives levels ranging from 0.1% to 0.3% of wild type, depending on the allele and the gene. The results indicate that the signals from mating pheromone might be mediated by the CDC70, CDC72 and CDC73 products. products.
Protoplast fusion of Aspergillus oryzae
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 216~220
As the bsic study about protoplast fusion of amylolytic fungus Aspergillus oryze and nonamyloytic sugar fermenter, Saccaromyces cerevsisae, the intraspecific protoplast fusion of A. oryzae was carried out and the properties of the obtained fusants were investigated. For protoplast fomation from mycellia of auxotrophs, Novozyme 234 as lytic enzyme was the most effective and optimal pH was determined to be pH 5.5-6.0. When the two types of protoplasts were treated with a fusogen including 30% PEG4000, they fused effectively and most of fusants were heterokaryons. Protoplasts aggregated with 30% PEG4000 after fusion treatment were observed by the microscope. Protoplast regeneration frequency was 1.46 to 13.8% and complementation frequency of fusion was 0.12 to 0.16. Fusant strains had a 1.5-fold DNA content compared to that of parent strain. And amylase activity was intermediate between those of parent strains.
Ethanol production from starch by protoplast fusion between aspergillus oryzae and saccharomyces cerevisiae
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 221~224
Amylolytic filamentous fungus, Aspergillus oryzae and nonamylolytic sugar fermentable yeast, Saccharomyces cerevisiae were fused by protoplast fusion in order to develope microorganisms having their intergrated function. Aminoacid auxotrophic properties were used as a genetic marker of protoplast fusion, and 35% PEG 4000 was used as a fusogenic agent. Complementation frequengy of fusion was
Obtained fusants showed the morphology of yeast strains, the amylase activity and the ethanol productivity. Among the properties of the fusants, morphology and prototrophic property were sustained stably but their ethanol productivity from starch was reduced. Although fusant strains had 0.5-fold ethanol productivity compared to that of S. cerevisiae in glucose medium, they produced ethanol from strach by direct fermentation.
Characterization of the Infectious Pancreatic Necrosis Virus (IPNV) isolated from Pan-Cultured Rainbow Trout in Korea
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 225~230
Infectious Pancreatic Necrosis Virus (IPNV) is one of the most important pathogens for inland fish farming and had been first reported in Korea from returning adult chum salmon (Oncorhynchus keta) at hatcheries on the east coast. During the past years, several viruses identified as IPNV were isolated not only from chum salmon, but also from gold fish (Carassius auratus), eel(Anguilla japonica), and rainbow trout (Salmo gairdneri). An isolate, coded DRT, from fingerlings of pan-cultured rainbow trout in Daechung Dam showed different serotype from three known reference serogroups of IPNV such as VR-299, Sp, and Ab. Antisera to three of these serotypes, however, partially neutralized the infectivity of this isolate. Anti-Sp type was rather effective than either anti-VR-299 or anti-Ab, implying DRT could be more closely related to Sp. DRT has been purified and its RNA genome segments were compated showing that the isolate does not belong to any of known serogroups even with some common antigenicity.
A new serotype confirmed by partial physical mapping of cDNA clones from the infectious pancreatic necrosis virus (IPNV) isolated in Korea
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 231~236
The larger segment of double stranded RNA genome from a new serotype of Infectious Pancreatic Necrosis Virus (IPNV), DRT, has been partially cloned at Sma I site in pUC19 and compared with the restriction maps of VR-299 and Sp. Restrction sites found in DRT was distinct and hence a new serotype. The cDNA clones of DRT were about 800, 850, and 1, 400 bp long each and do not share any common restiction site. It is not clear yet if there exist any overlapping sequences among them. This partial cloning, however, was sufficient for the comparison of restriction maps with the other serotypes.
Purification and Some Properties of an Intracellular Protease from Pseudomonas Carboxydovorans
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 237~244
A soluble intracellular protease from cells of Pseudomonas carboxydovorans, a carboxydobacterium, grown on nutrient broth was purified 68-fold in five steps to better than 95% homogeneity with a yield of 2.4% using azocasein as a substrate. The enzyme activity was not detected from cells grown on pyruvate, succinate, acetate, or CO as a sole source of carbon and energy. The molecular weight of the native enzyme was determined to be 53,000. Sodium dodecyl sulfate-gel electrophoresis revealed the purified enzyme a monomer. The enzyme was found to be a serine-type protease. The enzyme activity was inhibited completely by several divalent cations such as
. The enzyme was also inhibited by EGTA, but was stimulated by iodoacetamide. The optimal pH and temperature for the enzyme reaction were found to be 8.0 and
, respectively. The enzyme was inactive on CO dehydrogenase.
Extracellular proteases from bacillus licheniformis : partial purification and characterization
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 245~249
Extracellular proteases of Bacillus licheniformis were partially purified using ammonium sulfate fractionation and Sephadex G-75 gel filtration chromatography. The partial purification permited the weparation of two different protease activities, type I and type II. Protease type I is an enzyme with rather high protealytic activity toward dasein and was highly susceptible to organofluoride and EDTA inhibitions. It showed maximal proteolytic activity at pH 7.5 and was rapidly denatured at
. Protease type II is a protease with relatively lower proteolytic activity than the type I. It was also inhibited by 10mM of EDTA and 1mM of PMSF by 30 min incubation. The enzyme showed maximal activity at pH 8.0 and was denatured relatively slowly at
Isolation and Identification of Korean type Streptococcus mutans
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 250~258
S. mutans known as a causative causative agent of dental caries was isolated from a carious lesion of Korean in the present study. The physiological, biochemical characteristics and polysaccharide pattern of these isolates were compated with those of four laboratory strains ; AHT(a), FA-1F(b), LM7(e), and OMZ65(g). One hundred strains of oral streptococci were isolated from dental caries sites of Korean (one male and one female). Among these, 3 strains were identified as S. mutans. These strains were able to grow on selective media MS, MST, MSP, MSP1, MSB, MSBT and were stained dark pink when sprayed with solutions of mannitol and TTC. So, these strains were called strain 108, 110, and 120, respectively. Strain 108, 110, and 120 were bacitracin resistnt. As these strains contained particularly hippurate hydrolysis enzyme, they were distinguished from laboratory strains. Apart from laboratory strains, the strain 108 was not capable of fermenting lactose, the strain 110 was not able to ferment sorbitol, inulin, melibiose, raffinose and the strain120 was incapable of fermenting inulin, raffinose. All fractions of extracellular and ecll bound polysaccharide of the strain108, 110, and 120 were consisted of more glucan than fructan. Aside from laboratory strains, the isolated strains were composed of more water-insoluble glucans related adherence on solid surface than water-soluble. According to these results, the strain108, 110, and 120 had native characteristecs of S. mutans, but they were different from laboratory strains in some characteristics. Therefore, each of them was given a name to S. mutans KHC108, KHC110, and KHC120, respcetively.
A Study on the Adherence of Oral Streptococci to Saliva- or Protein-Coated Hydroxyapatite Beads
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 259~264
The adherence of
-labeled oral streptococcal cells to protein-coated hydroxyapatite (HA) beads was studied by a standard adherence assay. The adherence equilibrium for S. mutans 10449 occured in about 2 hrs. The cell numbers adhering to SHA was 50% less than those on bare HA. Sailva from different subjects had varying effect on bacterial adherence. The use of saliva adsorbed with homologouis bacteria decreased S. mutans adherence by 38% ; this indicates the presence of salivary agglutinin in acquired pellicle formed on HA. Animal sera and BSA decreased S. sanguis adherence. BSA concentration as high as 10mg/ml caused up to 87% adherence inhibition. The desorption experiment of adhered bacteria confirmed the previous reports that the adhesive sites on HA beads for S. mutans were different from those for S. sanguis and that S. mutans could enhance the adherence of S. sanguis but not vice versa.
Numerical Analysis of Bacterial Community in Cheonsu bay
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 265~271
Bacteria isolated from Cheonsu Bay at 4 seasons were analyzed by numerical taxonomic method. Results of 48 morphological, physiological and biochemical tests showed different adaptability of bacteria to temperature in consequence with sampling season and isolated bacteria were able to survive at various environmental conditions, Identification results revealed that Enterobacteriaceae, Aeromonas, Pseudomonas and Vibrio were dominant genera in geterotrophic bacterial community. For each season, Aeromonas was most dominant in spring and autumn, Pseudomonas and Enterobacteriaceae in summer and winter, respectively. Cluster ananlysis was performed and all vacteria were clustered into 29 phenetic groups. Seasonal characteristics were distict in each group. Different physiological characteristics and species compositions for each season contribute to the stability and diversity of environmental ecosystem.
Numerical taxonomy of 3-chlorobenzoate degrading bacteria isolated from Korean coastal waters
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 272~278
The bacteria utilizing 3-chlorobenzoate as a sole carbon and energy source were isolated by Most-Probable-Number technique and identified in Korean coastal waters. Pseudomonas, Moraxella and Flavobacterium were dominant genera and comprised 70.2% of total isolates. Forty-four biochemical, cultural, morphological and physiological testa were performed and average linding cluster analysis was conducted from the test results. Total 92.7% of isolates were clustered into 17 groups under the 80% similarity level. The degradation of 3-chlorobenzoate was performed by many heterogeneous bacteria and the species diversity of these bacterial group offers informations of the stability of bacterial communities in relation to carbon compound cycling in coastal enviromnents.
Isolation and characterization of sodium dodecylbenzenesulfonate(soft type)-degrading bacteria
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 279~284
Macroorganisms capable of utilizing sodium dodecylbenzenesulfonate(SDBS, soft type) as a sole carbon source were isolated from nature by using SDBS agar plate technique. Iwolated bacteria were examined primarily for biodegradation ability of SDBS, and followed by testing for resistance to several kinds of metal compounds and antibiotics. Among them(152 strains), one strain showed a excellent SDBS-degrading ability with a resistance to amipicillin and rifampicin was selected. This bacterium was identified as Klebsiella sp. and harbored two plasmids of about 4 and 5 kilobases. SDBS-degrading ability was lost when the plasmids were cured by mitomycin C. It was revealed that the degradation of SDBS was controlled by the plasmid DNA encoding genes. The two plasmids were stably maintained in Escherichia coli C600.
Effects of herbicide butachlor on Rhodospirillum rubrum KS-301
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 285~290
The biololgical effect of the preemergence rice field herbicide, butachlor(commercial name, Machete) on purple nonsulfur photosynthetic bacterium Rhodospirillum rubrum KS-301 has been studied under cultural conditions. Bacterial growth showed a tendency to decline according to the degree of the concentration of butachlor until
M and slmost stopped at
M. The growth inhibitory action at
M of butachlor was evident (4.2-18.7% inhibition of growth rate) but had little effect in nitrogen fixation. Conversely, there was a little enhancement effect(1%) in pyruvate, dinitrogen gas growing cultures. At concentration of
M, instead of spiral form, rod shapes were observed through phase contrast microscope and instead of vesicular intracytoplasmic membrane, irregular tubular forms were observed through electron microscope. Alkaline pH value slightly reversed tha inhibitory action of butachlor.
Disinfection effects of heat- and cold-treatment and UV-irradiation on campylobacter jejuni
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 291~296
Campylobacter jejuni was studied for its disinfection by heat-and cold-treatment and UV-irradiation. When C. jejuni was treated by heat, no viable cell was found after 10 min treatment at
, whereas small fraction of cell population was survived after 60 min treatment at
. When they were treated by cold temperature for 30 days, no cell was survived at -
but about 4 log of the cells were survived at both temperature of
. When the organisms were UV-irradiated, thier survival rates were proportionally varied to the distance of irradiation. The scanning electron microscopic studies of C. jejuni cells treated by the disinfecting agents revealed that shapes of thecells were deformed from spiral rod into spherical form. The heat-treated cells showed rough and damaged surface on the scanning electron micrographs. In the heat-treated cells, some proteins of high molecular weight appeared to become accumulated in the electrophoretic analysis. The DNAs extracted from the cells treated with the physical agents showed some differences in agarose gel electrophoresis, comparing those of normal cells.
Isolation and Characterization of Ultra-Violet and Gamma-radiation Resistant Bacteria from Natural Habitats
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 297~303
Attempts to isolate the naturally occurring ultra-violet resistant bacteria from environmental sources were made. The isolates, designated No.29, 100, and 107, among numbers of bacterial isolates revealed a remarkable resistance to UV ray, whose degree of resistance in dose/response kinetics was comparable to that of an endospore-former, Bacillus subtilis. In a range of 100-300
/min of UV irradiation, the isolates exhibited 500-1000 fold resistance compated with E. coli. The isolated appeared to possiss cell-bound pigment of organge or crimson-red. The isolate 29 is spherical in pairs or tetrads, whereas the isolates 100 and 107 are rod. All are Gram-gositive bacteria and seemed to be non-endospore-bearer. A number of biochemical studies pursued on the isolates suggested that they are quite different to each other. Electron microscopic examination and the physiological characters of the isolate 29 suggested that this UV resistant spherical bacterium might be one species of Deinococcus, probably Deinococus radiophilus. Since there is no documents on UV resistant, Gram-positive, non-sporeformer bacillus so far, the isolates 100 and 107 might be turned out as new kinds of UV resistant bacteria occurring in nature by further investigation.
Effect of simazine on nitrogen assimilation of rhodopseudomonas sphaeroides
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 304~309
The effects of simazine [2-chloro-4, 6-bis(methylamino)-s-triazine] on in vivo nitrogenase, glutamine synthetase, glutamate synthase, and glutamate dehydrogenase which are important in nitrogen assimilation of Rhodopseudomonas sphaeroides were investigated. Simazine inhibited the synthesis of nitrogenase and glutamine synthetase. The activity of glutamine synthetase in crude extracts of Rhodopseudomonas sphaeroides is less inhibited by simazine than that of glutamate synthase and glutamate dehydrogenase. These results suggest the possibility that simazine inhibits photosynthetic activity and so inhibits the synthesis of nitrogenase and glutamine synthetase in Rhodopseudomonas sphaeroides.
Studies on the microbiological assay method for tabtoxin produced in pseudomonas syringae pv. tabaci
The Korean Journal of Microbiology, volume 27, issue 3, 1989, Pages 310~315
Tabtoxin produced in Pseudomonas syringae pv. tabace irreversibly inhibits its known physiological target, glutamine synthetase so that causes wildfire disease on leaves of host plant. In this study, we examined a rapid and sensitive microbiological method for tabtoxin assay in several media. In minimal A agar medium nd minimal glucose agar medium, growth inhibition zone of Agrobacterium tumefaciens was larger than that of other indicator strain. However, mostly, growth inhibition zone of indicator strains on the minimal glucose agar medium was smaller than that of on the miniaml A agar medium. In complex agar medium, growth inhibithiton zone was not observed in all the tested indicator strains. Pseudomonas syringae pv. tabaci produced more tabtoxin according to the incubation time. When glutamine was added to the minimal glucose agar medium, growth inhkbition zone of Agrobacterium tumefaciens was reduced.