• Proceedings of the Microbiological Society of Korea Conference
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• 1991.04a
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• pp.237-242
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• 1991

In order to increase the production of glutathione by maximizing the expression of recombinant gsh plasmids, two genes responsible for the biosynthesis of glutathione were cloned. A gshI gene was cloned onto pBR322 plasmid as 3.6Kb PstI DNA fragment from E. coli K-12 chromosomal DNA. Also gshII gene was cloned onto pUC13 plasmid as 2.2Kb PstI-BamHI DNA fragment. In order to improve the glutathione producing activity more efficiently, various recombinant plasmids containing tandem repeated gshI genes or both genes in various copy number onto the same vector were constructed. E. coli cells harboring pGH501 plasmid (pUC8-gshI$\cdot$I$\cdot$II) showed the highest glutathione synthesizing activity. The conditions for glutathione production with an ATP-generating system such as acetate kinase reaction of E. coli cells or glycolytic pathway of yeast cells were examined using the E. coli cells harboring the pGH501 plasmid. When the acetate kinase reaction of E. coli cells was used as an ATP generating system, 20mM of L-csteine was converted into glutathione with a yield of $100\%$.

• Biomedical Science Letters
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• v.19 no.4
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• pp.310-314
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• 2013

CYP2E1 in the liver has been studied intensively because it is involved in the metabolic activation of xenobiotics. It is inducible by alcohol, so it has been suspected as the cause of cancer in the stomach and lung. The possible role of CYP2E1 has been suggested strongly as causing tissue damage in mice with renal failure. It was also suspected that 5'-flanking region of CYP2E1 gene might be involved with renal failure. So, we investigated polymorphism of restriction enzyme sites within CYP2E1 gene using the PCR-RFLP analysis. PstI and RsaI sites were located at 5'-flanking region and DraI site was located at intron 6. All three types (W/W, W/S, S/S) were observed for these enzymes although each incidence was somewhat different depending the enzyme sites. W/W was prominent for PstI whereas W/S was markedly high for RsaI. Overall, polymorphic incidence in patients was somewhat higher than normal population. This research should facilitate further investigation of CYP2E1 at genetic level as the direct cause of tissue damage in various organs.

• Korean Journal of Microbiology
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• v.38 no.1
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• pp.7-12
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• 2002

The intergenic spacer (IGS) region of the ribosomal DNA of species in Fusarium section Liseola was analyzed by amplification and subsequent digestion with several restriction enzymes. The length of the amplified IGS region was about 2.6 Kb in all strains except F.moniliforme 12 Which was about 2.9 Kb. The enzymes, EcoRI, HincII, SalI, HindIII, PstI and SmaI, digested the IGS region and nine haplotypes were identified among 11 strains. In the dendrogram based on PCR-RFLP of IGS region combined the results of section Liseola in this study and section Elegans in previous study, variation in the IGS appears to offer considerable potential to resolve intraspecific relationship as well as interspecies or intersection.

• Korean Journal of Microbiology
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• v.35 no.3
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• pp.205-210
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• 1999

16brio cholerne is an important pathogenic organism that causes dimhea in human beings. V ciaoleroe KNIH002 was isolated from patients suffering with dian.heal disease in Korea. From Southern hybridization using the amplified PCR product of 307 bp as a probe. which was obtained from PCR reaction using primer detecting cholera toxin gene, we have found that the c b gene located in 4.5-kb fragmenl double digested with Pstl and BgllI of the chromosome. Therefore, we made mini-libraries of the isolate using PstI and Bgm restriction endonuclease and pBluescript SKU(+) vector. As a result. we cloned 4.5-kb PstI-BglII fragment containing the c a gene encoding a cholera toxin from the constructed mini-libraries of V olzolerae KNlH002 by colony hybridization using the same probes. This recombinant plasmid was named pCTX75. E. coii XL1- Blue harboring pCTX75 showed the cytotoxicity on Chinese Hamster Ovary cells. From the sequencing of he cloned recombinant plasmid, we confinned that it has virulence gene cassette consisting of ace, zot, ctx.4 and cf"~B gene. The ace and zot genes were composed of 291 hp and 1.200 bp with ATG initiation codon and TGA lennination codon, respectively. Nucleotide sequence of the ace gene exhibited 100% identity with that of V cholera E7946 El Tor Ogawa strains. But, nucleolide and amino acid sequence comparison of the zot gene exhibited 99% and 98.8% identity with that of V cholerae 395 Classical Ogawa stram, respectively. Specially. the Ala-100, Ala-272 and Ala-281 sites of Zoi polypeptide presented in V choleme 395 Classical Ogawa strain are replaced by Val in V cholerae KNIH002.

• Proceedings of the Zoological Society Korea Conference
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• 1998.10b
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• pp.349.1-349
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• 1998

No Abstract, See Full Text

• Journal of Microbiology and Biotechnology
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• v.6 no.6
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• pp.386-390
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• 1996

The ${\beta}$-galactosidase gene from Lactococcus lactis subsp. lactis ATCC 7962 was cloned and its enzymatic properties were characterized, with a view to assessing its potential use as a selection marker in the food-grade cloning vector. Chromosomal DNA from L. lactis subsp. lactis 7962 was cleaved with PstI and ligated into pBR322 for transformation into Escherichia coli TGl. Transformants showing ${\beta}$-galactosidase activity possessed the pBR322 plasmid containing a 10 kilobase (kb) PstI fragment and this plasmid was named pCKL11. The cloned ${\beta}$-galactosidase gene came from the chromosomal DNA of L. lactis subsp. lactis 7962 was confirmed by Southern hybridization. A restriction map of pCKL11 was constructed from the cleavage of both pCKL11 and the purified 10kb insert fraqment. The. optimum pH of the ${\beta}$-galactosidase determined with the E. coli harboring the pCKL11 was 7.0. The optimum temperature was $50^{\circ}C$, while the pI of the enzyme was 7.4. These values were the same as those of the enzyme from the parent strain.

• Imaging Science in Dentistry
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• v.53 no.1
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• pp.1-9
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• 2023

Purpose: The aim of this study was to evaluate 3-dimensional cone-beam computed tomography (CBCT) images of alveolar bone changes in patients who underwent minimally invasive periodontal surgery-namely, the pinhole surgical technique (PST). Materials and Methods: Alveolar bone height was measured and compared on CBCT images of 254 teeth from 23 consecutive patients with Miller class I, II, or III recession who had undergone PST. No patient with active periodontal disease was selected for surgery. Two different methods were used to assess the alveolar bone changes postoperatively. In both methods, the distance between the apex of the tooth and the mid-buccal alveolar crestal bone on pre- and post-surgical CBCT studies was measured. Results: An average alveolar bone gain >0.5 mm following PST was identified using CBCT(P=0.05). None of the demographic variables, including sex, age, and time since surgery, had any significant effect on bone gain during follow-up, which ranged from 8 months to 3 years. Conclusion: PST appears to be a promising treatment modality for recession that results in stable clinical outcomes and may lead to some level of resolution on the bone level. More long-term studies must be done to evaluate the impact of this novel technique on bone remodeling and to assess sustained bone levels within a larger study population.

• Korean Journal of Microbiology
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• v.25 no.4
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• pp.282-289
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• 1987

The characteristics of the plasmid pKU10 isolated from Pseudomonas putida KU816 were investigated and its restriction map was constructed. The pKU10 plasmid was a small R plasmid carrying genes for resistance to ampicillin, tetracyclin, and chloramphenicol, and cured by treatment with mitomycin C. The molecular size of pKU10 was estimated to be 9.4Kb. Pseudomonas strains and E. coli cells could be transformed for antibiotic resistance characters specified by pKU10 plasmid DNA. By incompatibility test with other plasmids, pKU10 is grouped into IncP-1. EcoRI, XhoI, SalI, BglII, and SmaI cleaved pKU10 once, while PstI cleaved at two sites, and HindIII cleaved at six sites. The restriction map was constructed by partial and complete digestion of the purified plasmid DNA with single, double, or triple restriction enzymes. Thus, pKU10 is expected to be used for a cloning vector in Pseudomonas cells.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.6
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• pp.752-756
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• 2011

Polymorphism of the second exon of the caprine leukocyte antigen-DRB3 gene (CLA-$DRB3^*02$) was investigated in this study. The 285 bp PCR product of 258 individuals from 10 domestic goat breeds in Southwest China was digested with restriction endonucleases PstI and HaeIII and then genotyped. Three alleles and 4 restriction digestion profiles were distinguished by digestion of the PCR fragment by PstI, and 8 alleles and 13 genotypes by HaeIII. For HaeIII restriction enzyme sites, the Chi-square ($X^2$) test showed that all goat breeds in this study did not fit with the Hardy-Weinberg equilibrium (p<0.01 or p<0.05). The highly polymorphic nature of CLA-$DRB3^*02$ was demonstrated and the ranges of gene heterozygosity (He) and polymorphism information content (PIC) were 0.36-0.63 and 0.32-0.55, respectively. Clustering analysis showed that the 10 goat breeds clustered into two groups and Dazu Black goat had a close genetic relationship with Chengdu Grey, Jintang Black and Nanjiang Yellow goats.

• Journal of Life Science
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• v.8 no.3
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• pp.348-351
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• 1998

Common DNA fragments of the ${\beta}$-globin gene were observed from six races of the adult common carp: Hybrid-Yamato, Japanese wild type, Mirror, Suwa-Yamato, Scale German, and Saku-Yamato. Chromosomal DNAs isolated from the above six races were digested with restriction endonucleased EcoRI and PstI. The digested fragments were transferred onto nitrocellulose filter and hybridized with a probe of carp ${\beta}$-globin cDNA. Molecular sizes of the hybridized DNA fragments digested with EcoRI were 3.6Kb(Kilo base), 4.3Kb and 15Kb in Hybrid-Yamato, Japanese wild type, Mirror, Scale German and Saku-Yamato carp DNAs. In Scale German and Saku-Yamato carp DNAs, two and one more hybridized DNA fragments were observed, respectively. Molecular sizes of the hybridized DNA fragments digested with PstI were 2.2Kb, 6.5Kb, 7.8Kb and 9.2Kb in Hybrid-Yamato, 2.2Kb, 6.5Kb and 9.2Kb in Japanese wild type, 2.2Kb, 6.5Kb, 7.8Kb, and 13Kb in Mirror, 2,2Kb, 5,5Kb, 6.5Kb, 7.8Kb, 9.2Kb and13Kb in Scale German, and 2.2Kb, 5.5Kb, 6.5Kb, 9.2Kb and Saku-Yamato carp DNA. Therefore, depending on carps, three to six DNA fragments were hybridized with ${\beta}$-globin gene probe. Thus it indicated polymorphysm in the globin gene family of carp.