• Journal of Environmental Science International
• /
• v.1 no.2
• /
• pp.99-103
• /
• 1992

The bacteriophages lytic for Vibrio furnissi, Vibrio furniulis and Vibrio parahemolyticus were isolated from fish gills and shellfish. Nucleic acid of bacteriophage was prepared and restriction endonuclease profile was compared. All isolates contained deoxyribonucleic acid. V. fumissi bacteriophage from fish gills showed 2 bands with Bgl II, 1 with Pst, 3 with Hind III, 1 with Bm HI and 2 with EcoR I. V Puuialis phage represented 7 fragments with Bgl II, 1 with Pst, 4 with Hind III, and 2 with EcoR I. V parhemolyticn produced 13 sites with Hind III and 4 sites with EcoR I. The fragment types were varied depending on the phage isolation. All three phages were digested with Hind III and EcoR I with different sizes. V furnissi phage were digested with 5 different restriction enzymes. Key words: Bacteriophage, Vibrio furnissi, Vibrio fluvialis, Vibrio pnrahemolyticus, Deoxyribonucleic acid, Pst, Bam HI, Hind III, EcoR I, Bgl II.

• Environmental Sciences Bulletin of The Korean Environmental Sciences Society
• /
• v.1 no.2
• /
• pp.99-103
• /
• 1997

The bacteriophages lytic for Vibrio furnissi, Vibrio fluvialis and Vibrio parahaemolyticus were isolated from fish gills and shellfish. Nucleic acid of bacteriophages was prepared and restriction endonuclease profile was compared. All isolates contained deoxyribonucleic acid. V. furnissi bacteriophage from fish gills showed 2 bands with Bgl II, 1 with Pst, 3 with Hind III, I with Bam HI and 2 with EcoR 1. V fluvialis phage represented 7 fragments with Bgl II, 1 with Pst, 4 with Hind III, and 2 with EcoR 1. V. parahamolyticus produced 13 sites with Hind III and 4 sites with EcoR 1. The fragment types were varied depending on the phage isolation. All three phages were digested with Hind III and EcoR I with different sizes. V. furnissi phage were digested with 5 different restriction enzymes.

• Korean Journal of Microbiology
• /
• v.39 no.3
• /
• pp.187-191
• /
• 2003

In an attempt to isolate myxobacteria from soil samples, we isolated swarm and fruiting body forming bacteria that have bacteriolytic activity on Coli-spot agar plate. For the classification of myxobacteria, 16S rDNA RFLP patterns were analyzed. Amplified 16S rDNAs of myxobacteria type strains (Family I, II, III and IV), negative control strains and soil-isolates were restricted with HaeIII, EcoRI and EcoRV, respectively. We found that the soil-isolates belongs to myxobacteria Family I, II, III.

• Korean Journal of Veterinary Research
• /
• v.33 no.1
• /
• pp.43-51
• /
• 1993

Mitochondrial DNA(mt DNA) of Mammalian is the circular one which the 16.5K base pairs and show the maternal inheritance. Evolutional speed of nucleotide sequence is very fast. So that polymorphic analysis of mt DNA provide the useful informations to investigate the genetic relations of interspecies. Authors trials were focussed to compare with the polymorphic differences of mitochondrial DNA between Jindo and Japanese mongrel dogs. DNA was extracted from bloods of 21 head of Jindo dogs and 20 head of Japanese dogs and isolated using 10 kinds of restriction endonucleases(Apa I, BamH I, Bgl II, EcoR I, EcoR V, Hinc II. Hind III, Pst I, Sty I, Xba I) and then separated by the agarose gel electrophoresis. After sourthern blotting hybridization was completed using the mtDNA of Japanese mongrel dogs as a probe. Autoradiography was used to compare the polymorphism of mtDNA both dogs. The results obtained were as follows; 1. mt DNA of Jindo dog showed polymorphism resulting cleavage with four kinds of restriction endonuclease, Apa I, EcoR V, Hinc II, Sty I. While in the Japanese mongrel dogs observed the polymorphism in the five kinds of restriction endonuclease supplemented with EcoR I. 2. Compared with both dogs the frequency differences of DNA polymorphism were recognized in the specific restriction endonuclease Apa I. Consequently in the restriction endonuclease Apa I both dogs classified with three types as A, B, C however in the Jindo dogs frequency of C type was 71.5 percent but in Japanese mongrel dogs observed 45 percent in the A type. 3. DNA polymorphism obtained from the use of five kinds of restriction endonuclease were classified with seven types. In Jindo dogs frequency was highest in the type 6 as 71.4 percent but in the Japanese mongrel dogs showed 35 percent in the type 5. 4. Genetic distances calculated by NEI method showed 0.0089 in Jindo dogs and was 0.0094 in the Japanese mongrel dogs.

• Microbiology and Biotechnology Letters
• /
• v.22 no.6
• /
• pp.593-598
• /
• 1994

A bacterium KL-57 which exhibited biosurfactant activity was isolated. This bacterium was identified as Bacillus subtilis. The biosurfactant-producing gene of B. subtilis KL-57 was cloned into R subtilis MI113 by using plasmid pTB523. The plasmid DNA from the clone was found to carry a 18 kb PstI insert. The biosurfactant-producing gene was cleaved into 4 fragments by SmaI, 3 fragments by PvulI or EcoRl, 4 fragments by PvulI and EcoRI double digestion, 5 fragments by AccI, and 2 fragments by KpnI, HindIII or BamHI. By subcloning the 18 kb Pstl insert, a 2.3 kb EcoRl fragment conferred the biosurfactant producing activity on B. subtilis cells. The 2.3 kb had one HindIII cleave site. But Two fragments, which corresponds HindIII/EcoRl termini, exhibited no biosurfactant activity.

• Korean Journal of Microbiology
• /
• v.21 no.2
• /
• pp.95-102
• /
• 1983

The penicillin G acylase(pga) gene was cloned in the vector plasmid pKM $300(Ar^r,\;Tc^r,\;6.33kb)$ for the study of the structure and expression of the pga gene. This recombinant plasmid pPAKS-1 DNA(24.5 Kb) was cleaved into 2 fragments by restriction enzyme Eco R1.1fragment by BamH1, 4fragments by Hind III, and 2 fragments by Pst I. The pga gene was located on the Eco R1.Hind III-C fragement of pPAKS-1. The recombinant plasmids pPAKS-1 and pPAKS-2, in which the Hind III-B and Hind III-D fragments pPAKS-1 are deleted, are characterized. The results are summarized as follows : 1. Doubling times of bacterial strain bearing pPAKS-1 and pPAKS-2 are 90 and 60 minutes, respectively. 2. pPAKS-1 and pPAKS-2 are present at about 16-32 and 70 copies per cell, respectively, are 0.66 and 5.5 units, respectively, which represent 2-fold and 20-fold higher enzyme 4. pPAKS-1 is very unstable, but pPAKS-2 is stable.

• The Korean Journal of Food And Nutrition
• /
• v.9 no.2
• /
• pp.123-136
• /
• 1996

IPSKCDNA gene(1.8 kbp) encoding rat brain IP3K enzyme contained Not I restric site in open reading frame. The Not I sequence, GCGGCCGC, was converted to GCAGCCGC by site-directed mutagenesis. The mutated IP3KcDNA was digested with EcoR I and ligated with EcoR I-restricted psp72·Not2 vector. The resulting psp72 · Not2-IP3KCDNA was digested with the Not I restriction enzyme and then subcloned into the Not I -digested PZIP · NeoSV(X) mammalian expression vector. The PZIP · NeoSV(X) -IPSKCDNA was transfected into CCL39 hamster lung fibroblast cells. The efficiency of the expressed IPSKCDNA gene was significantly higher than expected generally, not only a mean 5-fold increase in the amount of enzyme, but also 16-fold increase in enzyme activity from tractsfected CCL39 cells by the method of Western blot using anti-lP3K antibodies. Both distribution of IPSK in various rat tissues and biochemical properties were discussed.

• Journal of Environmental Science International
• /
• v.5 no.5
• /
• pp.605-610
• /
• 1996

The bacteriophage from the fresh oyster, Crassostrea Virginica which is specific to the marine bacterium was isolated and characterized. Among the foci different vibrio species and the five different serotypes of Vibrio parahaemolyticus host strains tested, only two strains of the parahaemolyticus possessing K17, K52 antigens were highly sensitive to the phage. The size of the isolated plaque was 0.4mm and the electron microscopic head size of the isolated phage was about 67 nm long and 83 nm wide. PFU/ml was 1.25$\times$ $10^{11}$. The phase was sensitive to chloroform but resistant to acetone or methanol. The assay of the isolated phase nucleic acid was deoxyribonucleic acid. The restriction enzyme pattern showed 14 fragment from Hind III and 4 fragments from Eco R I. Two different antigenic groups showed-similar restriction enzyme patterns.

• Microbiology and Biotechnology Letters
• /
• v.17 no.5
• /
• pp.447-453
• /
• 1989

In order to study the transfer of antibiotics resistance genes of the genetically cloned bacteria in water environments, DK1 strain, which is resistant to kanamycin (Km), chloramphenicol (Cm), streptomycin (Sm), and sulfadiazine (Su), was selected from the Gram-negative bacterial isolates from wastewater. One of 4 plasmids harboured in the DK1 strain was found to possess Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ gene and be about 68 kb in size, and it was designated as pDK101. The plasmid of pDK101 was also found to have 16, 32, and 6 restriction sites for EcoRI. .PstI, and SalI, respectively. From the digestion fragments of pDK101 plasmid and pKT230 used as a vector by EcoRI restriction endonuclease, pDT309 and pDT529 were constructed as chimeric plasmids which possess Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ gene and are 30.9 and 52.9 kb in size, respectively. When the chimeric plasmids were trasformed into E. coli C600 or E. coli HB101, transformants of DKC601, DKC602, DKH102, and DKH103 were obtained as cloned bacterial cells. The Km$^{${\gamma}$}$Cm$^{${\gamma}$}$ genes were well expressed in those cloned cells and the chimeric plasmids were clearly detected in the cloned cells of DKC601 and DKH103.

• Environmental Mutagens and Carcinogens
• /
• v.22 no.3
• /
• pp.199-204
• /
• 2002

It has been known that Ganoderma lucidum and Lentinus edodes have anticancer activity and immune enhancing activity. These two mushrooms were grown in liquid culture and harvested. From these mycelia, DNA was isolated and EtBr-CsCl density gradient ultracentrifugation was performed to purify it further. Then mitochondrial DNA was isolated by bisbenzimide-CsCl density ultracentrifugaton. Mitochondrial DNA of Ganoderma lucidum was digested by restriction enzymes, EcoR I, Hind Ⅲ, and Pst I, then electrophoresed. It showed 12, 22, 4 fragments. Mitochondrial DNA of Lentinus edodes was digested by EcoR I. Electric pattern showed 6 fragments. 4 fragments had appeared by Pst 1 digested mitochondrial DNA. Hind ill couldn't digest mitochondrial DNA of Lentinus edodes. Mitochondrial DNA of fusants was isolated to compare to those of parents. The results showed that fusant P₂S₄has new, recombined mitochondrial DNA. But P₂S₄had the same DNA that Ganoderma lucidum had.