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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 29, Issue 6 - Dec 1991
Volume 29, Issue 5 - Nov 1991
Volume 29, Issue 4 - Sep 1991
Volume 29, Issue 3 - Jul 1991
Volume 29, Issue 2 - May 1991
Volume 29, Issue 1 - Mar 1991
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Cloning and Expression of Human Immunodeficiency Virus-1 Epitopes in Escherichia coli
The Korean Journal of Microbiology, volume 29, issue 1, 1991, Pages 1~7
Human immunodeficiency virus type 1 (HIV-1) causes a deadly infectious disease, Acquired Immunodeficiency Syndrome (ADIS). As a first step to develop a reliable and fast diagnostic procedure for HIV-1 infection, we cloned various immunodominant epitopes of HIV-1 in bacterial expression vectors containing tac or trp promoter. While the protein level of direct expression of gp160 was low, trp E fused gp120, gp41 and p17-p24 were produced at high levels (15-30% of total bacterial proteins) in E. coli. Since gp120 and gp41 contain relatively conserved regions which can react with antibodies in the plasma from most of HIV-1 infected individuals, these expression clones were used for large preparations of HIV-1 antigens.
Construction of Interspecific Hybrids detween Aspergillus spp. by Nuclear transfer
The Korean Journal of Microbiology, volume 29, issue 1, 1991, Pages 8~15
Interspecific hybrids between the ASpergillus spp., A. awamori, A. usamii and A. oryzae, were obtained by nuclear transfer technique. Nuclei isolated from an auxotrophic mutant strain were transferred into the protoplasts of a recipient strain of different species. The frequency of interspecific hybrid formation by nuclear transfer was
In contrast, no interspecific hybrid was isolated by protoplast fusion. Among the hybrids tested, 10 strains showed increased activity of some or all components of cellulases, xylanases and amylase up to more than two times. Isozyme pattern of the hybrids were analyzed by polyacrylamide gel electrophoresis and isoelectric focusing followed by activity staining, which showed that some of the hybrids have isozyme patterns unidentical to either of the two parents. By measuring the DNA contents and the sizes ofthe conidia, the karyotypes of the hybrids were estimated to be aneuploid near to haploid, diploid or triploid. It was concluded that the unclear transfer technique is much more efficient in the formation of interspecific hybrids than protoplast fusion and is very useful for the improvement of Aspergillus strains.
Molecular Cloning, Chromosomal Integration and Expression of the Homoserine Kinase gene THR1 of Saccharomyces cerevisiae
The Korean Journal of Microbiology, volume 29, issue 1, 1991, Pages 16~24
The yeast gene THR1 encodes the homoserine kinase (EC 126.96.36.199: HKase) which catalyses the first step of the threonine specific arm at the end of the common pathway for methionine and threonine biosynthesis. A recombinant plasmid pMC3 (12.6 kilobase pairs, vector YCp50) has been cloned into E. coli HB101 from a yeast genomic library through its complementing activity of a thr1 mutation in a yeast recipient strain M39-1D. When subcloned into pMC32 (8.6kbp, vector YRp7) and pMC35 (8.3 kbp, vector YIp5), the HindIII fragment (2.7 kbp) of pMC3 insery was positive in the thrI complementing activity in both yeast and E. coli auxotrophic strains. The linearized pMC35 was introduced into the original recipient yeast strain and the mitotically stable chromosomal integrant was identified among the transformants. Through the tetrad analysis, the integration site of the pMC35 was localized to the region of THR1 structural gene at an expected genetic distance of approximately 11.1 cM from the ARG4 locus on the right arm of the yeast chromosome VIII. When episomically introduced into the auxotrophic cells and cultured in Thr omission liquid medium, the cloned gene overexpressed the HKase in the order of thirteen to fifteenfold, as compared with a wildtype. HKase levels are repressed by addition of threonine at the amount of 300 mg/l and 1, 190 mg/l for pMC32 and pMC3, respectively. Data from genetic analysis and HKase response thus support that the cloned HindIII yeast DNA fragment contains the yeast thr1 structural gene, along with necessary regulatory components for control of its proper expression.
Antiviral Activity of Papaverine and Nucleoside Analogs on the Human Cytomegalovirus Infection
The Korean Journal of Microbiology, volume 29, issue 1, 1991, Pages 25~33
Antiviral activities of papaverine and nucleoside analogs, 9-[(1,3-dihydroxy-2-propoxy)methyl]guanine (DHPG) and acyclovir, against human cytomegalovirus (HCMV) infection were compared in vitro. Papaverine and DHPG were effective in reducing infectious HCMV yields with
(effective dose 50: the concentraion at which 50% of virus yields was obtained) of approximately 1.02 and
, respectively; while acyclovir was less effective with an
The relative cytotoxicity of these drugs was evaluated under the same conditions used to measure infectious HCMV yields. Papaverine and DHPG demonstrated little cellular toxicity as measured by their effect on the viability of confluent cells at concentrations in the range of those demonstrating potent inhibition of HCMV replication. Similarly, protein synthesis was largely unaffected by these drugs in stationary mock-infected cells as measured by the incorporation of isotopically labelled amino acids. In contrast, cellular DNA synthesis was invariably reduced in the presence of either drug. HCMV-specific DNA synthesis was also strongly inhibited by papaverine and DHPG.
cis-Diamminedichloroplatinum (II) (CDDP) Inhibits Bluetongue Virus (BTV) Core Associated Transcriptase Activity
;Manning, JaRue S.;
The Korean Journal of Microbiology, volume 29, issue 1, 1991, Pages 34~39
The BTV core associated transcriptase activity, assayed by acid precipitable counts, was reduced to an undetectable level after treat the core with .
CDDP. When the RNA transcripts prepared from the CDDP treated BTV core were analysed on agaroseurea gel, it was observed that the band intensity of the large size RNA was reduced while the band intensity of the small size RNA was enhanced. Northern blot analysis showed that much of the small size RNAs appeared to be prematurely terminated transcripts. These results suggest that CDDP adduction to the template RNA blocks chain elongation process of the virion bound transcriptase that is ultimately responsible for the inactivation of BTV infectivity.
Identification of Malonate-specific Enzymes, Malonyl-CoA Synthetase and Malonamidase, in Rhizobia
The Korean Journal of Microbiology, volume 29, issue 1, 1991, Pages 40~48
Two malonate-specific enzymes, malonyl-CoA synthetase and malonamidase, were found in free-living cultures of Rhizobium japonicum, Rhizobium meliloti, and Rhizobium trifolii, that infect plant roots where contain a high concentration of malonate. Malonyl-CoA synthetase catalyzes the formation of malonyl-CoA, AMP, and PPi directly from malonate, coenzyme A, and ATP in the presence of
Malonamidase is a novel enzyme that catalyzes hydrolysis and malonyl transfer of malonamate, and forms malonohydroxamate from malonate and hydroxylamine. Both enzymes are highly specific for malonate. These results show that Rhizobia have enzymes able to metabolize malonate and suggest that malonate may be used in symbiotic carbon and nitrogen metabolism.
Synthesis and thermotolerance of heat shock proteins in campylobacter jejuni
The Korean Journal of Microbiology, volume 29, issue 1, 1991, Pages 49~55
The heat shock responses of Campylobacter jejuni were studied by examination of their survival rates and synthesis of heat shocd proteins. When C. jejuni cells were treated at the sublethal temperatures of 48.deg.C for 30 minutes, most of the cells maintained their viabilities and synthesized the heat shock proteins of 90, 73, and 66 kD in molecular weight. By the method of two-dimensional electrophoresis, the heat shock proteins of C. jejuni were identified to be Hsp90, Hsp73, and Hsp66. During the heat shock at 48.deg.C, the heat shock proteins were induced from about 5 minutes after the heat shock treatment. Their synthesis was continued upto 30 minutes, but remarkably retarded after 50 minutes. When C. jejune cells were heat shocked at 51.deg.C for 30 minutes, the survival rates of the cells were decreased by about
fold and synthesis of heat shock proteins and normal proteins was also generally retarded. The cells exposed to 55.deg.C for 30 minutes died off by more than
cells and the new protein synthesis was not observed. But when C. jejuni cells were heat-shocked at the sublethal temperature of 48.deg.C for 15 to 20 minutes and then were exposed at the lethal temperature of 55.deg.C for 30 minutes, their viabilities were higher than those exposed at 55.deg.C for 30 minutes without pre-heat shock at 48.deg.C. Therefore, the heat shock proteins synthesized at the sublethal temperature of 48.deg.C in C. jejuni were thought to be responsible for thermotolerance. However, when C. jejuni cells heat-shocked at various ranges of sublethal and lethal temperatures were placed back to the optimum temperature of 42.deg.C, the multiplication patterns of the cells pretreated at different temperatures were not much different each other.
Identification and Characteristics of Extreme Halophilic bacteria Isolated from a Saltern in Korea
The Korean Journal of Microbiology, volume 29, issue 1, 1991, Pages 56~62
Extremely halophilic bacteria isolated from salterns at Mado, Kyunggido, Korea, were identified and investigated on their salt requirements. The results have shown that six strains were identified to be belonged to the genus Halobacterium and three strains identified as the fenus Halococcus. Among them, the optimal NaCl concentration for growth of Halobacterium sp. EH10 was at 4.2M and no growth occurs below 2.0M NaCl. The strain, EH10, is nonmotile and showed acid production from glucose, fructose and maltose while H. salinarum is motile and does not produce acid from any carbohydrates. On the other hand, the strain EH10 does not utilize readily glucose while a number of sugars are readily utilized for growth with acid production by H. saccharovorum. Thus, the isolate, EH10, was classified into the genus Halobacterium and could be a novel species of the genus by its main morphological and physiological features including G+C content. The optimal temperature for growth of the isolate, EH10, was 50.deg.C. But this strain did not grow when NaCl was replaced with KCl.
Genetic Characteristics of Arsenic Compounds-Resistant Bacteria Isolated from Stream Water
The Korean Journal of Microbiology, volume 29, issue 1, 1991, Pages 63~68
Several arsenic compound-resistant bacteria were isolated from Jung-Rang stream. The isolates, D-3, D-12, and D-14 were characterized phenotypically and genetically, and identified as Serratia liquefaciens, Klebsiella oxytoca, and Klebsiella pneumoniae, respectively. A plasmid of 67kb was found in Klebsiella oxytoca D-12 and designated as pMH12. Transfer of this plasmid from D-12 to E. coli HB101 was occurred, and the resulting transconjugant strains expressed the same level of heavy metal resistance as the donor strain. The physical presence of this plasmid in transconjugant was detected with agarose gel electrophoresis. Arsenite-sensitive derivatives of isolate D-12 were obtained with Mitomycin C treatment which cured pMH12. Antibiotics and heavy metal resistances were also examined to be used as a proper marker for the isolates in gene cloning. Isolate D-12 has resistance to several heavy metal ions such as
Also, all the other arsenite resistant isolates showed resistance to several heavy metal ions and antibiotics.
Chromosome Number in Several Species of the Genus Fusarium
The Korean Journal of Microbiology, volume 29, issue 1, 1991, Pages 69~73
The chromosome of Fusarium species during the vegetatve nuclear divisions in hyphae were observed by use of HCl-Giemsa technique on light microscope. The haploid chromosome number of Fusarium anthophilum 7472 was n=7, n=6 in F. anthophilum 7481 and n=6 in F. oxysporum 7500. The haploid chromosome number was 7 in F. napiforme 6129 and F. napiforme 6144. Those of F. caucasicum F. caucasicum ATCC 18791 and F. aquaeductuum ATCC 15612 were n=5. F. coeruleum ATCC 20088 was n=6, n=8 in F. camptoceras ATCC 16065 and n=7 in F. sambucinum NRRL 13451. From these results and previous papers, it may be concluded that the basic haploid chromosome number of the genus Fusarium is n=4.