• Journal of Ginseng Research
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• v.42 no.1
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• pp.42-49
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• 2018

Background: Ginsenoside F1 has been described to possess skin-whitening effects on humans. We aimed to synthesize a new ginsenoside derivative from F1 and investigate its cytotoxicity and melanogenesis inhibitory activity in B16BL6 cells using recombinant glycosyltransferase enzyme. Glycosylation has the advantage of synthesizing rare chemical compounds from common compounds with great ease. Methods: UDP-glycosyltransferase (BSGT1) gene from Bacillus subtilis was selected for cloning. The recombinant glycosyltransferase enzyme was purified, characterized, and utilized to enzymatically transform F1 into its derivative. The new product was characterized by NMR techniques and evaluated by MTT, melanin count, and tyrosinase inhibition assay. Results: The new derivative was identified as (20S)-$3{\beta},6{\alpha},12{\beta}$,20-tetrahydroxydammar-24-ene-20-O-${\beta}$-D-glucopyranosyl-3-O-${\beta}$-D-glucopyranoside(ginsenoside Ia), which possesses an additional glucose linked into the C-3 position of substrate F1. Ia had been previously reported; however, no in vitro biological activity was further examined. This study focused on the mass production of arduous ginsenoside Ia from accessible F1 and its inhibitory effect of melanogenesis in B16BL6 cells. Ia showed greater inhibition of melanin and tyrosinase at $100{\mu}mol/L$ than F1 and arbutin. These results suggested that Ia decreased cellular melanin synthesis in B16BL6 cells through downregulation of tyrosinase activity. Conclusion: To our knowledge, this is the first study to report on the mass production of rare ginsenoside Ia from F1 using recombinant UDP-glycosyltransferase isolated from B. subtillis and its superior melanogenesis inhibitory activity in B16BL6 cells as compared to its precursor. In brief, ginsenoside Ia can be applied for further study in cosmetics.

• Journal of Microbiology and Biotechnology
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• v.4 no.1
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• pp.20-23
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• 1994

The polymerase chain reaction was used to study the genes inducible under stress from the heavy metal cadmium. Schizosaccharomyces pombe, grown in the presence or absence of sublethal concentration of cadmium, was isolated to purify the total RNAs. The Induced RNA Random Fishing (IRRF) method in which random oligonucleotides were used as primers was applied to the identification of cadmium-induced gene expressions. A PCR-DNA product of 400-bp was cloned and sequenced. Computer analysis showed that this DNA has no homology with any known DNA sequences in GenBank or EMBL databases. The induction of this gene was confirmed by Northern blot analysis of total RNAs isolated from both cadmium-treated and untreated yeast cells.

• International Journal of Industrial Entomology and Biomaterials
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• v.15 no.2
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• pp.137-143
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• 2007

Lactate dehydrogenase (LDH) is a ubiquitous enzyme that plays a significant role in the clinical diagnosis of pathologic processes. Discovery of the LDH (BmLDH) gene in B. mori may shed light on its role in the biology of Lepidoptera species, and afford further understanding of the function of the enzyme. In this study, we used the bioinformatics tools to identify LDH gene in B. mori. Sequence analysis showed that BmLDH cDNA contains a 996 bp open reading frame, encoding 331 AA proteins, with seven introns. Compared with hHLDH (human heart LDH), BmLDH contained the same key active sites. Domain search and protein fold recognition analyses provide compelling evidences that the deduced protein is a LDH. Using the computer program MEGA3, we conducted a search for homologs of BmLDH among many eukaryotic species and confirmed that the BmLDH was conserved in all organisms investigated. This gene has been registered in GenBank under the accession number EU000385.

• Korean Journal of Microbiology
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• v.46 no.2
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• pp.162-168
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• 2010

The sequence diversity of mcyA gene involved in synthesis of microcystins was analyzed in Microcystis spp. isolated from the Korean reservoirs and in the environmental samples taken from the Daechung, Chungju, Yongdam, Soyang, and Euam Reservoirs at the cyanobacterial blooming season. It was estimated that the sequences of mcyA gene in the isolated Microcystis spp. were much conserved when compared with those in GenBank database. A few kinds of clones were dominant in the investigated environmental samples, occupying 87 to 100% of total clones. No mcyA sequences originated from Anabaena spp. or Planktothrix spp. was found. These results indicated that microcystins are produced mainly by Microcystis spp. and the sequences of mcyA genes are much conserved in the investigated Korean reservoirs.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2005.04a
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• pp.59-60
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• 2005

• Journal of Microbiology and Biotechnology
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• v.15 no.5
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• pp.1120-1124
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• 2005

Pseudomonas putida KCTC 1639 was newly identified as a potential producer of biodegradable medium chain length polyhydroxyalkanoates. It exhibited a carbon assimilation pattern quite different from other known P. putida strains, but a more similar pattern with P. oleovorans, which assimilates the carbon sources mainly through ${\beta}$-oxidation rather than the fatty acid biosynthesis pathway. The PHA synthesis gene cluster from P. putida KCTC1639 was composed of two gene loci; the PHA synthase gene locus and granule-associated gene locus, which were cloned and deposited in the GenBank under accession numbers AY286491 and AY750858 as a new nucleotide sequence, respectively. The molecular structure and amino acid homology of the new gene cluster were compared with those from Pseudomonas species, including other P. putida strains and P. oleovorans, and a higher than $90\%$ homology was observed.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.16
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• pp.7061-7069
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• 2015

Background: During the past decades, the incidence and mortality rate of stomach cancer has demonstrated a great decrease in the world, but it is still one of the most common and fatal cancers especially among men worldwide, including Iran. The MYC proto-oncogene, which is located at 8q24.1, regulates 15% of genes and is activated in 20% of all human tumors. MYC amplification and overexpression of its protein product has been reported in 15-30% of gastric neoplasias. The aim of this investigation was to find the relative efficacy of CISH (chromogenic in situ hybridization) or IHC (immunohistochemistry) in diagnosis and prognosis of gastric cancer, as well as the relationship of amplification and expression of C-MYC gene with patient survival. Materials and Methods: In this cross-sectional study, 102 samples of gastric cancer were collected from patients who had undergone primary surgical resection at the Cancer Institute Hospital, Tehran University of Medical Sciences, from July 2009 to March 2014. All samples were randomly selected from those who were diagnosed with gastric adenocarcinomas. CISH and IHC methods were performed on all of them. Results: Patients were classified into two groups. The first consisted of stage I and II cases, and the second of stage III and IV. Survival tests for both groups was carried out with referrnce to CISH test reults. Group II (stage III & IV) with CISH+ featured lower survival than those with CISH- (p=0.233), but group I (stage I & II) patients demonstrated no significant variation with CISH+ or CISH- (p=0.630). Kaplan-Meier for both groups was carried out with IHC test findings and showed similar results. This data revealed that both diffuse and intestinal types of gastric cancer occurred significantly more in men than women. Our data also showed that CISH+ patients (43%) were more frequent in comparison with IHC+ patients (14.7%). Conclusions: For planning treatment of gastric cancer patients, by focusing on expanding tumors, which is the greatest concern of the surgeons and patients, CISH is a better and more feasible test than IHC, in regard to sensitivity and specificity. Therefore, CISH can be used as a feasible test for tumor growth and prognosis in stage III and IV lesions. This study also indicated that C-MYC amplification in gastric cancer is correlated with survival in advanced stages.

• Animal Systematics, Evolution and Diversity
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• v.18 no.1
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• pp.13-21
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• 2002

To add genetic information to the conservation efforts on grey coral (Naemorhedus caudatus) in Korea, we investigated the pattern of mitochondrial cytochrome b gene sequence (606 bp) of six specimens from two localities in Korea. The corresponding sequences of N. caudatus in China obtained from GenBank were also used. The nucleotide Tamura-Nei distances between each of four haplotypes of N. caudatus in Korea and the haplotype of N. caudatus in China varied from 0.0650 to 0.0803: N. caudatus revealed high level of sequence diversity in Bovidae. In N. caudatus in Korea, the distances among three haplotypes at Yanggu were 0.0151 to 0.0185, and it suggests that the genetic diversity of Yanggu population was decreased in low level. Moreover, the distances between each of three haplotypes at Yanggu and one haplotype at Samcheok were 0.0343 to 0.0489. It indicates that habitat isolation caused the continuous increase of genetic distance with geographic distance in N. caudatus, and various conservation plans for mitigating the loss of genetic diversity in Korea have to be in immediate action. To clarify the taxonomic status of N. caudatus, the sequence (276 bp) of N. goral available from GenBank were also utilized, and n goral was not distinct from N. caudatus. It suggests that they may be conspecific, but further analyses with additional specimens of two species are necessary.

• The Plant Pathology Journal
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• v.19 no.3
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• pp.159-161
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• 2003

The coat protein (CP) gene of Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was selected for the design of virus-specific primers for amplification and molecular detection of the virus in cultivated apple. A combined assay of reverse transcription and polymerase chain reaction (RT-PCR) was performed with a single pair of ApMV-specific primers and crude nucleic acid extracts from virus-infected apple for rapid detection of the virus. The PCR product was verified by restriction mapping analysis and by sequence determination. The lowest concentration of template viral RNA required for detection was 100 fg. This indicates that the RT-PCR for detection of the virus is a 10$^3$times more sensitive, reproducible and time-saving method than the enzyme-linked immunosorbent assay. The specificity of the primers was verified using other unrelated viral RNAs. No PCR product was observed when Cucumber mosaic virus (Cucumovirus) or a crude extract of healthy apple was used as a template in RT-PCR with the same primers. The PCR product (669 bp) of the CP gene of the virus was cloned into the plasmid vector and result-ant recombinant (pAPCP1) was selected for molecule of apple transformation to breed virus-resistant transgenic apple plants as the next step. This method can be useful for early stage screening of in vitro plantlet and genetic resources of resistant cultivar of apple plants.

• The Plant Pathology Journal
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• v.23 no.3
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• pp.151-154
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• 2007

Phytoplasma was identified from leaf lettuce (Lactuca sativa) cultivated in commercial green-house in Korea. Diseased leaf lettuce revealed proliferation of shoots, and yellowing and shrinking of leaves (lettuce proliferation-K). Polymerase chain reaction (PCR) with universal primer pair P1/P6, and aster yellows (AY) specific primer pair R16F1/R1 amplified 1.5kb and 1.1kb length of DNA fragments, respectively. Nucleotide sequences of 16S rRNA gene were determined (Gen Bank accession no EF489024). Phylogenetic analysis of 16S rDNA showed the closest relationship with AY phytoplasma (GenBank accession no. AY389822 and AY389826), indicating that lettuce proliferation-K is a member of AY. Phytoplasma bodies were detected in phloem sieve tubes of diseased lettuce by transmission electron microscopy. The structures had round or pleomorphic shapes with a diameter of 130-300nm. Phylogenetic analysis of 16S rRNA gene, microscopic observation of phytoplasma bodies and symptomatology indicated that lettuce proliferation-K is caused by phytoplasma in the AY group. This is the first report of phytoplasma disease in lettuce in Korea.