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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 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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The Structure and The Reason for Nuclear Accumulation of Poly A(-) Spliced SV40 RNA
The Korean Journal of Microbiology, volume 27, issue 1, 1989, Pages 1~9
The locations of 5' ends as well as the splicing pattern of viral poly A(-) 19S RNA from monkey cells infected with SV40 were determined by a modification of primer extension method. The 5' end of this RNA mapped at the major cap site at nucleotide residue 325, used most frequently by SV40 late RNAs. The intron from nt.373 to nt.558 was removed as the ordinary cytoplasmic poly A(+) 19S RNA. The 3'end of this RNA was very heterogeneous and distributed over 1 kb upstream of polyadenylation site, as determined by S1 nuclease mapping. The reason for this normally initiated and spliced RNA to accumulate in the nucleus was investigated. In order to test whether the presence of unused 3' splice region on this RNA caused such subcellular distribution, cells were transfected with SV40 mutant KNA containing deletion around 3' splice site. The RNA deleted of 3' splice region accumulated mainly in the cytoplasm. This accumulation did not result from the increased stability of the RNA due to the deletion, since the wild type and mutant RNAs exhibited similar half lives after chase with actinomycin D. Therefore it is likely that the 19S spliced RNA is hindered from being transported into the cytoplasm due to some pre-splicing complexes formed at the unused 3' splice site.
Transforming Capacity of the Plasmid Containing SV40 Promoter in NIH3T3 Fibroblast Cells
The Korean Journal of Microbiology, volume 27, issue 1, 1989, Pages 10~15
The plasmid pKOneo, containing SV40 transcriptional promoter, has been used in the mouse tumorigenicity assay for oncogene studies. This assay employs a cotransfection of NIG3T3 fibroblast cells with the desired DNA and the plasmid pKOneo. This oncogene assay, however, has been speculated due to the SV40 transcriptional promoter in the plasmid pKOneo. This research was designed to investigate if the plasmid pKOneo alone is capable of transforming NiH3T3 fibroblast cells. The NIH3T3 subclones were established after the NIH3T3 cells were transfected with the plasmid pKOneo alone. The estabilished NIH3T3 subclones, containing the exogeneous plasmid pKOneo in their chromosomes, were examined for their expression of transformation-associated parameters. The results indicate that this plasmid pKOneo alone has positive effects on transformation of NIH3T3 cells after integration into cellular chromosomes.
Effect of escherichia coli plasmid DNA sequences on plasmid replication in yeast
The Korean Journal of Microbiology, volume 27, issue 1, 1989, Pages 16~20
The effect of E. coli plasmid DNA sequences contained by chimeric vectors on plasmid replication was investigated. We constructed YRp7- or 2.
m circle-based plasmids containing E. coli plasmid DNA sequences and those not containing it. By examining their maintenance in yeast, we showed that plasmid without E. coli plasmid DNA sdquences was nore stable and presented higher copy number, and espressed higher level of hepatitis B viral surface antigen as a foreign gene. This result suggested that E. coli plasmid DNA sequences within chimeric plasmid somehow inhibited plasmid replication in yeast.
Cloning of nif Gene Cluster from Klebsiella pneumoniae and Expression in Escherichia coli
The Korean Journal of Microbiology, volume 27, issue 1, 1989, Pages 21~26
The chromosomal DNA of Klebsiella penumoniae KNF1 was partially digested with HindIII. pBR322 was completely digested with same enzyme with additional alkaline phosphatase treatment. Both DNA samples were ligated and transformed into E. coli KO60. Single coloneis of the transformed cells were isolated on N0free agar media. These colonies were ampicillin-resistant and tetracycline-sensitive. When the plasmids in transformants were cured, nitrogen-fixing activities were lost. Therefore, these transformants harbored recombinant plasmid including nif genes of K. pneumoniae. Nitrogenase activity of transformant was higher than or the same as that of K. pneumoniae KNF1.
Identification of hybride from intra- and interspecific protoplast fusion in trichoderma by electrophoretic patterns of enzymes
The Korean Journal of Microbiology, volume 27, issue 1, 1989, Pages 27~34
In order to evaluate the applicability of enzyme electrophoresis for the identification of intra/interspecific hybride obtained by the protoplast fusion in Trichoderma, soluble proteins, intracellular soluble enzymes and extracellular
-glucosidase were analyzed by polyacrylamide gel electrophorsis. As the results, patterns of soluble protein, and isozyme patterns of peroxidase, malate dehydrogenase, and
-glucosidase in hydrids were defferent from those in parental and wild type strains. Therefore, it was established that the analysis of protein pattern by electrophoresis could be applied to isolate and identify the hybrids from the protoplast fusion.
H-NMR spectroscopic evidence on the glycosidic linkages of the transglycosylated products of low-molecular-weight
-D-glucosidase from trichoderma koningii
The Korean Journal of Microbiology, volume 27, issue 1, 1989, Pages 35~42
The mode of transglycosylation reaction observed during the action of low-molecular-weigh
-D-glucoside glucohydrolase, EC18.104.22.168) purified from Trichoderma koningii ATCC 26113 was investigated using
-NMR spectroscopy. The enzyme was purified by the series of procedures including ammonium sulfate precipitation, and fractionations by column chromatographies on Bio-Gel P-150, DEAE-Sephadex A-50, and SP-Sephadex C-50. The final purification was performed by the band eluation after preparative polyacrylamide gel electrophoresis. The enzyme showed its molecular size of 78,000 through the analysis of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and its isoelectric point of 5.80 through the analysis of analytical isoelectric focusing. The H-1 proton resonances were analyzed. After the reaction of the enzyme with cellobiose, the reaction products were separated by high performance liquid chromatography using refractive index detector. H-1 resonances of the products were consisted with those of gentiobiose [
-D-glucopyranosyl--(1,6)-D-glucopyranose], and cellotriose [
-D-glucopyranosyl]-(1,4)-D-glucopyranose] with minor resonances of sophorose [
Protease released during germination of dictyostelium discoideum spores
The Korean Journal of Microbiology, volume 27, issue 1, 1989, Pages 43~47
Characteristics and roles of protease released during the germination of Dictyostelium discoideum spores were investigated. When geat activated, the spores germinated, progressively releasing the protease into the extracellular medium. The protease activity exhibited high at pH 2.5. When cyclogeximide was added to culture, complete germination (emergence) and protease release were stopped. Addition of purified nonspecific protease to culture speeded up germination. These results suggest that excreted protease may play a role in removal of the spore wall.
Improvement of an hydroxyapatite bead adherence assay for streptococcus sanguis
The Korean Journal of Microbiology, volume 27, issue 1, 1989, Pages 48~55
The purpose of the present investigation was to improve several procedures being used in the adherence assay of Streptococcus sanguis cells to hydroxyapatite (HA) beads and to study the effect of the beads on the counting of radioactivity. The standard adhere assay involved the adherence of radiolabeled bacteria to 40mg of HA beads. The beads were mixed with ['H]thymidine-labeled bacterial cells and incubated for 60 minutes at room temperature. Unadsorbed cells were removed, the beads with adsorbed cells were dried, and the radioactivity was monitered in a scintillation spectrometer. The 30 seconds sonication of cells in a form of long chains appeared to be adequate for obtaining mostly singlet or doublet cells. Unlike the counting of S. sanguis cell suspension, bacterial cells adhered to HA or saliva-coated HA(SHA) required smaller volume (2.5ml) of scintillaton fluid for better counting. Eighteen percent quenching of counts could be attributed to the beads. Among 3 procedures commonly used to equilibrate the beads for adherence assay, no differences were found in their effectiveness. The HA beads on which the bacteria remained attached in scintillant during the counting were found to be the source of sample self-absorption representing 34.5% of the total radioactivity counts resulting from the beads dissolved in HCl solution.
Antagonistic activity of Streptomyces apecies against Fusarium solani causing ginseng root rot
The Korean Journal of Microbiology, volume 27, issue 1, 1989, Pages 56~62
Antagonistic effects of Streptomyces species aganinst Fusarium solani causing ginseng root rot were investigated in terms of chitinase activity and growth inhibition in vitro. Among 131 isolates of streptomycetes obtained from ginseng cultivating soil, 9 isolates producing large clear zone around the colony on a chitin agar medium were selected for further study. All 9 isolates produced chitinase in a range from 0.10 to 0.38 U lysing cells of F. solani and inhibited germination of the conidia. In the ten-fold condentrated culture filtrate of S. alboniger ST59 and S. roseolilacinus ST129, the number of conidia of F. solane was reduced to about 20% of original count within 14 days. When S. alboniger ST59 and F. solani were grown simultaneously in the mineral saly medium, chitinase activity increased with incubation period, whereas mycelial volume of F. solani decreased. In a chitin added mineral salt medium, chitinase activity increased during the first four days and maintained steady level until the 8th day, and increased thereafter. S. alboniger ST59 lysed mycelia, conidia and even chlamydospores of F. solani. It is probable that the antagonistic activity of this streptomycete against F. solani is the lysis of fungal cell wall by streptomycete producing chitinase affected by antifungal substances.
Isolation and Characterization of a Pink-Pigmented Facultative Methylotrophic Bacterium
The Korean Journal of Microbiology, volume 27, issue 1, 1989, Pages 63~69
A pink-pigmented facultative methylotrophic bacterium, Methylobacterium sp. strain SY1, was isolated from soil through methanol-enrichment culture technique. The isolate was gram-negative, slightly curved rod, and motile by a single polarly inserted flagellum. The colony was smooth, bright pink, and slimy. The guanine plus cytosine content of the KNA was 66%. The cell was obigately aerobic and exhibited both catalase and oxidase activities. Carotenoid pigment and poly-
-hydroxybutyrate were present. It was found to have three kinds of plasmid with molecular weights 45,000, 38,500 and 23,000. Growth with methanol(0.5%) was fast (
=6.5h) and was optimal at
and at pH 7.0. The isolate could grow on several sugars, organic acids, amino acids, amines, and alcohols in addition to the methanol. Methanol was found to be assimilated through the serine pathway.
Bioconversion of progesterone by immobilized aspergillus phoenicis
The Korean Journal of Microbiology, volume 27, issue 1, 1989, Pages 70~76
Progestrone bioconversion by immobilized Aspergillus phoenicis was studied. Progesterone was converted into 11
-hydroxyprogesterone and 3-minor byproducts. Whole cells of A. phoenicis were immobillized by enreappment with calcium-alginate, K-carrageenan, or polyacrylamide. Of these materials tested, cell immobilized in
-alginate gels showed the highest activity for 11
-hydroxylation of progesterone. In the case of mycelia immobilized in
-alginate, futher progressing hydroxylation of 11
-hydroxyprogesterone was greatly reduced. Spores of A. phoenicis which were immobillized with
-alginate and germinatedin situ for 25 hours showed higher 11
-hydroxylase activity than those of entrapped whole mycelia and maintained initial enzyme activity for all 8 times of repeated use. After 16 times of reuse, the activity was declined 30% or more. When culture media and
were introduced into the reaction media, the activity of the immobilized mycelia which had been lowered due to many times of reuse was effectively reactivated.