• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.311-317
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• 2006

Using PCR amplification, we cloned a cellulase gene (ce/H) from the Bacillus subtilis AH18 which has plant growth-promoting activity and antagonistic ability against pepper blight caused by Phytophthora capsici. The 1.6 kb PCR fragment contained the full sequence of the cellulase gene and the 1,582 bp gene deduced a 508 amino acid sequence. Similarity search in protein database revealed that the cellulase of B. subtilis AH18 was more than 98% homologous in the amino acid sequence to those of several major Bacillus spp. The ce/H was expressed in E. coli under an IPTG inducible lac promoter on the vector, had apparent molecular weight of about 55 kDa upon CMC-SDS-PAGE analysis. Partially purified cellulase had not only cellulolytic activity toward carboxymethyl-cellulose (CMC) but also insoluble cellulose, such as Avicel and filter paper (Whatman No. 1). In addition, the cellulase could degrade a fungal cell wall of Phytophthora capsici. The optimum pH and temperature of the ce/H coded cellulase were determined to be pH 5.0 and $50^{\circ}C$. The enzyme activity was activated by $AgNO_3$ or $CoCl_2$. However its activity was Inhibited by $HgC1_2$. The enzyme activity was activated by hydroxy urea or sodium azide and inhibited by CDTA or EDTA. The results indicate that the cellulase gene, ce/H is an antifungal mechanism of B. subtilis AH18 against phytophthora blight disease in red-pepper.

• Microbiology and Biotechnology Letters
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• v.48 no.3
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• pp.287-295
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• 2020

Lactobacillus zymae GU240 was previously isolated from Kimchi, a Korean fermented vegetable, as a strong GABA producer. The strain showed β-galactosidase (β-Gal) activity on MRS agar plates with X-gal. When growth and β-Gal activities of GU240 were measured using MRS (glucose, 2%, w/v) and MRSL (lactose, 2%, w/v) broths, cells were found to grow slowly in MRSL, and the β-Gal activity (36 units at 4 h) was lower than that of cells grown in MRS (94 units at 16 h). The highest OD₆₀₀ value of the culture in MRS was 1.6 at 24 h at 37℃, whereas that of the culture in MRSL was 0.6 at 16 h. β-Gal activity of the culture in MRS reached the maximum (95.6 u/ml) at 16 h, decreased thereafter, and was not detected at 48 h. β-Gal activity for culture in MRSL reached its highest (36 u/ml) at 4 h and decreased gradually, but some activity (11.05 u/ml) still remained at 72 h. The structural gene encoding β-Gal in L. zymae GU240 was cloned as a 3.1 kb fragment, and DNA sequencing confirmed the presence of complete lacLM genes. lacLM genes from L. zymae GU240 showed 98-99% homologies in nucleotide sequences with other lacLM genes from L. brevis. Reverse transcription (RT)-PCR confirmed the operon structure of lacLM. The results indicated that L. zymae GU240 might be in the process of losing the ability to grow rapidly on lactose-containing medium, such as milk, due to adaptations to plant environments, including kimchi.

• The Korean Journal of Mycology
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• v.45 no.4
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• pp.386-393
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• 2017

Onion downy mildew, caused by Peronospora destructor, is a major disease in onion cultivation areas in Korea. The causal fungi were collected and analyzed based on sequence similarity and molecular phylogenetic relationships of multi-gene sequences, including the internal transcribed spacer (ITS) region. All isolates from Changnyeong-gun, Hamyang-gun, and Hapcheon-gun in Gyeongnam province, and Muan-gun, Haenam-gun, and Sinan-gun in Jeonnam province were identical in the four types of gene sequences, indicating they were genetically the same strains. In this study, a PCR method was developed based on the ITS gene sequences to amplify the specific DNA fragment for P. destructor only. The detection limit of was total genomic DNA of the P. destructor and the plant $0.7ng/{\mu}L$. Therefore, the developed PCR method could be used to detect P. destructor effectively from symptomless onion leaves.

• The Journal of Korean Obstetrics and Gynecology
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• v.24 no.3
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• pp.14-27
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• 2011

Objectives: This study was designed to investigate the effects of Acanthopanacis Cortex Radicis extract(ACRE) on the apoptosis in HeLa cell and MCF-7 cell. Methods: After treatment with various concentration of ACRE, cell growth was evaluated in HeLa cell and MCF-7 cell. Hoechst 33342 staining was performed to estimate DNA fragment effect of ACRE on the apoptosis in HeLa cell and MCF-7 cell. Annexin V/PI apoptosis assay was used to estimate the effects of ACRE on the early apoptosis in HeLa cell and MCF-7 cell. RT-PCR was used to estimate the apoptosis gene expression effect of ACRE on Hela cell MCF-7 cell. Results: Under $0.1mg/m\ell$ of ACRE, cytotoxic effect was not found per NIH3T3 cell. The viability of HeLa cell and MCF-7 cells was significantly decreased ACRE ($100{\mu}g/m\ell$) in HeLa cell and MCF-7 cell, ACRE ($50{\mu}g/m\ell$) in HeLa cell 3 days after treatment, in MCF-7 cell 1&3 days after treatment (p<0.01). DNA fragmentation was observed 3 days after treatment of cl of ACRE on HeLa cell and MCF-7 cell. In Annexin V/PI apoptosis assay, after treatment of $100{\mu}g/m\ell$ of ACRE, the early apoptotic cell increased both in HeLa cell and MCF-7 cell. In RT-PCR analysis, after treatment of $100{\mu}g/m\ell$ of ACRE, bcl-2 were decreased and bax, caspase-3 were increased both in HeLa cell and MCF-7 cell. Conclusions: ACRE appears to have considerable activity on the apoptosis in HeLa cell and MCF-7 cell.

• The Journal of Korean Obstetrics and Gynecology
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• v.24 no.3
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• pp.1-13
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• 2011

Objectives: This study was designed to investigate the analysis of apoptosis by Scirpi Tuber in Hela cell and MCF-7 cell. Methods: For cytotoxic effect of Scirpi Tuber extract, Scirpi Tuber extract were cultured on NIH3T3 cell in vitro. After treatment with various concentration of Scirpi Tuber, cell growth was evaluated in Hela cell and MCF-7 cell. Hoechst 33342 staining was performed to estimate DNA fragment effect of Scirpi Tuber on the apoptosis in Hela cell and MCF-7 cell. Annexin V/PI apoptosis assay was used to estimate the effects of Scirpi Tuber on the early apoptosis in Hela cell MCF-7 cell. All the stained cells were analyzed by a FACS. RT-PCR was used to estimate the apoptosis gene expression effect of Scirpi Tuber extract on Hela cell and MCF-7 cell. Results: Cytotoxic effect of Scirpi Tuber extract was not found on per NIH3T3 cell. The viability of Hela cell was significantly decreased Scirpi Tuber (500, $1000{\mu}g/m\ell$) in Hela cell 1day, 3day and 5days after treatment (p<0.01). The viability of MCF-7 cell was significantly decresed Scirpi Tuber ($1000{\mu}g/m\ell$) in MCF-7 cell (p<0.01), Scirpi Tuber ($500{\mu}g/m\ell$) in MCF-7 cell only 3days after treatment (p<0.01). In RT-PCR analysis, after treatment of $100{\mu}g/m\ell$ of ACR extract, BCL-2 were decreased and BAX, caspase-3 were increased both in Hela cell and MCF-7 cell. DNA fragmentation was observed the Scirpi Tuber on Hela cell and MCF-7 cell. As time goes on DNA fragmentation incresed. In Annexin V/PI apoptosis assay, after treatment of $1mg/m\ell$ of Scirpi Tuber, the early apoptotic cell increased both in Hela cell and MCF-7 cell. As time goes on apoptotic cell increased. Conclusion: Scirpi Tuber appears to have considerable activity on the apoptosis in Hela cell and MCF-7 cell.

• Journal of Korea Water Resources Association
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• v.41 no.4
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• pp.423-432
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• 2008

The local migration or movement behavior of fishes in streams are related to feeding, spawning, growing, dispersing, and refuging. The pale chub (Zacco platypus) is a dominant species that migrates locally and inhabits in river and stream in Korea. However, dams, weirs, culverts and other regulatory structures are physical barriers that limit fish movement and fragment habits and populations. If main stream and off-channel habitats are connected with culverts, they would restrict the small fish as pale chub movement due to the high flow velocities and low depths. But in Korea, there is no experimental study to evaluate the swimming performance of species in Korea. Therefore, it is difficult to proposed that design guidelines for pass fishes through culverts. The purpose of this experimental study is to evaluate the swimming performance of pale chubs. A series of swimming performance test has been used in both of the fixed velocity and the incremental velocity methods in an experimental flume. As a result, the critical swimming speed for pale chub(body length 8.9 cm) was found to be about 0.7 m/s. Therefore, the flow velocity for culvert design in the low flow condition should not be exceed the its swimming ability, especially 0.7 m/s for pale chubs(body length 8.9 cm). And the minimum depth for culvert design in the low flow condition should not be lower than the fish body height add a dorsal fin height.

• Animal Systematics, Evolution and Diversity
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• v.15 no.2
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• pp.153-164
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• 1999

One hundred and eleven samples of six subspecies of striped field mouse, Apodemus agrarius Pallas from Korea, China and Russia, were used for the analysis of mitochondrial DNA(mtDNA) fragment patterns resulted from the digestion with eight restriction enzymes by blot hybridization technique. All 32 fragments, nine mtDNA haplotypes, and four major subgroups with the mean divergence value of 0.896 to 1.150% were revealed. In summary, three forms are recognized: [I, subspecies chejuensis (Chejudo island, Korea)], [II, subspecies pallescens (southwestern Korea), coreae (central Korea), and septentrionalis (Russia)], and [III, subspecies manchuricus (northeastern China) and pallidior (northern China)], although some samples of subspecies coreae are somewhat different from almost all samples of six subspecies, and some samples of subspecies pallidior are similar with all samples of subspecies septentrionalis to form same haplotype. It is confirmed that A. agrarius chejuensis is a distinct subspecies, that subspecies coreae (including pallescens) is also a distinct subspecies, that subspecies manchuricus and pallidior are synonyms of subspecies ningpoensis, and that subspecies septentrionalis is a synonym of subspecies ningpoensis, and that subspecies septentrionalis is a synonym of subspecies agrarius. Moreover, it seems that A. agrarius shows constant karyotype, minimal variation in mtDNA genotype, and considerable divergence in morphometric characters, although further analyses with additional samples of A. agrarius in Eurasia will be necessary to determine the degree of variation of these taxonomic characters and to clarify subspecies classification as well.

• Korean Journal of Microbiology
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• v.32 no.1
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• pp.47-52
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• 1994

The cells of Campylobacter jejuni heat-shocked at 48${\circ}C$ for 30 min synthesized the heat shock proteins of HSP90, HSP66 and HSP60. Those heat shock proteins were found to correspond to the heat shock proteins of HSP87, HSP66 (DnaK), and HSP58 (GroEL) of E. coli, respectively. By Southern blot analysis of the chromosomal DNAs of C. jejuni with groESL and dnaK genes of E. coli as DNA probes, the heat shock genes of C. jejuni which are homologous to the E. coli groESL and dnaK genes were found to exist in the chromosomal DNA. The genomic libraries of C. jejuni were constructed with the cosmid vector pWE15 and the groEL gene of C. jejuni were cloned in E. coli B178 groEL44 temperature senstive mutant. The hybrid plasmid (pLC1) was inserted with the DNA fragment (about 5.7kb in size) containing the groEL gene. E. coli groEL44 mutant cell transformed with the pLC1 could grow at 42${\circ}C$ by synthesizing the HSP60 of C. jejuni and regained the susceptibility to the ${\lambda}$ vir phage by expression of the groEL gene in the cloned cells. These indicated that the groEL products of C. jejuni had chaperon effects by synthesizing the heat shock proteins in the cloned cells of E. coli.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.2
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• pp.443-449
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• 2015

Background: Myeloproliferative disorders (MPDs) are clonal hematologic malignancies originating at the level of the pluripotent hematopoietic stem cell. Matrix metalloproteases (MMPs) are proteolytic enzymes that contribute to all stages of malignancy progression. Genetic variants in the MMP genes may influence the biological function of these enzymes and change their role in carcinogenesis and progression. To our knowledge, this is the first investigation of associations between the -735 C/T and -1562 C/T polymorphisms in the MMP2 and MMP9 genes, respectively, and the risk of essential thrombocytosis (ET), and polycythemia vera (PV). Materials and Methods: The case-control study included JAK2V617F mutation positive 102 ET and PV patients and 111 controls. Polymorphisms were determined by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and electrophoresis. Results: No statistically significant differences were detected between patient (ET+PV) and control groups regarding genotype distribution for MMP2 gene-735 C/T and MMP9 gene -1562 C/T polymorphisms and C/T allele frequency (p>0.050). Statistically borderline significance was observed between PV and control groups regarding genotype distribution for the MMP9 gene -1562 C/T polymorphism (p=0.050, OR=2.26, 95%Cl=0.99-5.16). Conclusions: Consequently this study supported that CC genotype of MMP9 gene -1562 C/T polymorphism may be related with PV even if with borderline significance.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.18
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• pp.8227-8233
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• 2016

Background: To investigate polymorphisms in heat shock proteins A1B and A1L (HOM) and associated risk of oesophageal carcinoma in Northeast India. Materials and Methods: The study includes oesophageal cancer (ECA) patients attending general outpatient department (OPD) and endoscopic unit of Gauhati Medical College. Patients were diagnosed based on endoscopic and histopathological findings. Genomic DNA was typed for HSPA1B1267 and HSPA1L2437 SNPs using the polymerase chain reaction with restriction fragment length polymorphisms. Results: A total of 78 cases and 100 age-sex matched healthy controls were included in the study with a male: female ratio of 5:3 and a mean age of $61.4{\pm}8.5years$. Clinico-pathological evaluation showed 84% had squamous cell carcinoma and 16% were adenocarcinoma. Dysphagia grades 4 (43.5%) and 5 (37.1%) were observed by endoscopic and hispathological evaluation. The frequency of genomic variation of A1B from wild type A/A to heterozygous A/G and mutant G/G showed a positive association [chi sq=19.9, p=<0.05] and the allelic frequency also showed a significant correlation [chi sq=10.3, with cases vs. controls, OR=0.32, $p{\leq}0.05$]. The genomic variation of A1L from wild T/T to heterozygous T/C and mutant C/C were found positively associated [chi sq=7.02, p<0.05] with development of ECA. While analyzing the allelic frequency, there was no significant association [chi sq=3.19, OR=0.49, p=0.07]. Among all the risk factors, betel quid [OR=9.79, Chi square=35.0, p<0.05], tobacco [OR=2.95, chi square=10.6, p<0.05], smoking [OR=3.23, chi square=10.1, p<0.05] demonstrated significant differences between consumers vs. non consumers regarding EC development. Alcohol did not show any significant association [OR=1.34, chi square=0.69, p=0.4] independently. Conclusions: It can be concluded that the present study provides marked evidence that polymorphisms of HSP70 A1B and HSP70 A1L genes are associated with the development of ECA in a population in Northeast India, A1B having a stronger influence. Betel quid consumption was found to be a highly significant risk factor, followed by smoking and tobacco chewing. Although alcohol was not a potent risk factor independently, alcohol consumption along with tobacco, smoking and betel nut was found to contribute to development of ECA.