• Ocean and Polar Research
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• 제43권1호
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• pp.31-43
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• 2021

To understand the temporal variation of prokaryotic communities in a temperate coastal area, prokaryotic abundance, activity, and community composition were investigated every week for over a year at a coastal monitoring station of Yeong-do, Busan. The prokaryotic abundances fluctuated about 10 times, ranging from 2.0 to 20.1 × 10⁵ cells mL^-1 and tended to be high in spring when phytoplankton bloom occurred. The prokaryotic thymidine incorporation rates (TTI) varied in a low range between 0.2 and 11.5 pmol L^-1 h^-1 in winter. However, in summer, TTI were increased up to a range of 8.3 to 17.4 pmol L^-1 h^-1, showing an increasing pattern in summer. During the study period, Alphaproteobacteria was the most dominant class for most of the year, followed by Flavobacteria. While the seasonal variation of prokaryotic composition was not apparent at the class level, many prokaryotic species showed a distinct temporal or seasonal variation for the year. In the coastal site, prokaryotic biomass and activity did not show significant correlations with temperature and chlorophyll-a, which are well known to regulate prokaryotic growth in marine environments, suggesting that the study area may be affected by diverse sources of organic matter for their growth.

• Environmental Analysis Health and Toxicology
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• 제23권1호
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• pp.33-39
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• 2008

The genotoxicity of mercury compounds have been investigated with a variety of genetic endpoints in prokaryotic and eukaryotic cells. The mercury ions are positively charged and easily form complexes with DNA by binding with negatively charged centers to cause mutagenesis. Further, the mercury ions can react with sulfhydryl (-SH) groups of proteins associated with DNA replication and alter genetic information. Another mechanism by which mercury damages DNA molecule is via its probable involvement of reactive oxygen species (ROS) and induces DNA strand breaks. In order to investigate whether the ROS production was induced by mercury, we performed ROS assay. As the result, the ROS production was significantly increased when it grows dose-dependently and time-dependently. We compared mercury alone-treated group and mercury co-treated with Vitamin C or glutathione group. As the result, the ROS production induced by mercury was decreased by Vitamin C and glutathione. Co-treated with Vitamin C and glutathione group was the most effective to lowering ROS production induced by mercury.

• Asian-Australasian Journal of Animal Sciences
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• 제20권6호
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• pp.827-831
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• 2007

Myostatin is a member of the transforming growth factor-${\beta}$(TGF-${\beta}$ super-family. It acts as a negative regulator for skeletal muscle growth. Myostatin mutations are characterized by a visible, generalized increase in muscle mass in double muscled cattle breeds. To understand the biochemistry and physiology of the Chinese Yellow bovine myostatin gene, we report here for the first time expression of the gene in Escherichia coli (E. coli). Primers of the myostatin gene of Chinese Yellow Cattle were designed on the basis of the reported bovine myostatin mRNA sequence (Gen-Bank Accession No. NM005259) and optimized for E. coli codon usage. XhoI and EcoRI restriction enzyme sites were incorporated in the primers, and then cloning vector and expression vector were constructed in a different host bacterium. The expressed protein had a molecule mass of about 16 kDa as determined by SDS-PAGE under reducing conditions. The expressed protein reacted specifically with myostatin monoclonal antibody on immunoblots. Our studies should lead to the investigation of the differences in myostatin genes of various cattle and could benefit human health and food animal agriculture.

• Journal of Microbiology and Biotechnology
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• 제12권2호
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• pp.306-311
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• 2002

O-linked N-acetylglucosamine (O-GlcNAc) is an abundant posttranslationally modified compound in eukaryotic cells. Human O-GlcNAc transferase (OGT) was produced as a maltose binding protein (MBP) fusion protein, which showed significant catalytic activity to modify recombinant Sp1, transcription factor. To facilitate the production of O-GlcNAc modified proteins, instead of using the tedious in vitro glycosylation reaction or expression in eukaryotic cells, a MBP-fusion OGT expression vector (pACYC184-MBPOGT) was constructed using pACYC184 plasmid, which could coexist with general prokaryotic expression vectors containing ColE1 origin. By cotransforming pACYC184-MBPOGT and pGEX-2T vectors into Escherichia coli BL21, intracellular O- GlcNAcylated proteins could be obtained by a simple purification procedure. It is expected that this may be a useful tool for production of O-GlcNAc modified proteins.

• Journal of Microbiology
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• 제42권3호
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• pp.200-204
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• 2004

Retron is a prokaryotic genetic element that produces multicopy single-stranded DNA covalently linked to RNA (msDNA) by a reverse transcriptase. It was found that cells producing a large amount of msDNA, rather than those that did not, showed a higher rate of mutation. In order to understand the molecular mechanism connecting msDNA production to the high mutation rate the protein patterns were compared by two dimensional gel electrophoresis. Ten proteins were found to be differentially expressed at levels more than three fold greater in cells with than without msDNA, nine of which were identified by MALDI TOF MS. Eight of the nine identified proteins were repressed in msDNA-producing cells and, surprisingly, most were proteins functioning in the dissimilation of various carbon sources. One protein was induced four fold greater in the msDNA producing cells and was identified as a 30S ribosomal protein S2 involved in the regulation of translation. The molecular mechanism underlying the elevated mutation in msDNA-producing cell still remains elusive.

• Journal of Microbiology and Biotechnology
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• 제14권6호
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• pp.1280-1285
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• 2004

Retron is a prokaryotic genetic element, producing a short single-stranded DNA covalently linked to RNA (msDNA-RNA) by a reverse transcriptase (RT). In retron EC83, msDNA is further processed at between the 4th and the $5^{th}$ nucleotides, leaving a 79 nucleotide-long single-stranded DNA as a final product. To investigate this site-specific cleavage in msDNA synthesis, we purified the RT protein of retron EC83. Initially, RT ORF was cloned under the tac promoter, but the expression was very poor largely because of poor translation. In order to facilitate translation, the nucleotide sequence for the first nine amino acids was randomized with synonymous codons. This change of downstream sequence of translational initiation codon greatly affected the efficiency of translation. We could isolate clones which greatly increased RT production, and their sequences were compared to those of the low producers. The overproduced protein was purified and was shown to have RT activity.

• 산업기술연구
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• 제28권B호
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• pp.59-63
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• 2008

Rapid production of therapeutic proteins such as angiostatin and endostatin angiogenic inhibititors has been highly demanded for cancer treatment. In this regard, recombinant human angiostatin and endostatin were successfully expressed as soluble forms by maltose binding protein (MBP)-mediated fusion expression in Escherichia coli. PCR amplified, angiostatin and endostatin genes from human placenta cDNA library were inserted into an expression vector pMAL-c2e to construct prokaryotic expression vectors, pMAL-c2e/AS and pMAL-c2e/ES, respectively. Recombinant angiostatin and endostatin were efficiently expressed in E. coli origami (DE3) after IPTG induction and protein expression were confirmed by SDS-PAGE analyses. The expressed recombinant proteins were purified near homogenity using an amylose affinty column chromatography. In contrast that previous E. coli expressions were all insoluble, our results first time demonstrated that MBP fused human angiostatin and endostatin were soluble in E. coli.

• 환경생물
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• 제18권4호
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• pp.441-446
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• 2000

Trichoderma harzianum is an aggressive causal agent of green mold disease on mushroom cultivation. Some bacterial strains isolated, from oyster mushroom compost in Wonju, were found to have in vitro antifungal activity against Trichoderma harzianum ATCC 6385, 6504, and our isolates Trichoderma spp. Y and G. Further in vitro antifungal studies on several strains of phytopathogenic fungi showed that all of 12 phytopathogenic fungal strains were significantly inhibited by the isolated antifungal bacteria in Petri dishes. Of these, KATB 99121 showed the broadest inhibiting effect and displayed as negative coagulase, negative sulfide production and rod shape. KATB 99121 was resistant to ampicillin, chlorampenicol, and kanamycin. Identification of isolates was determined by Biolog GN system, and KATB 99121 was identified as Photobacterium logei because of 96 probability, 0.65 similarity, and 4.97 disturbance. With electron microscopy, thin section of KATB 99121 strain revealed typical rod-like shaped cell (0.6-0.8${\mu}{\textrm}{m}$$\times$1.5-2.0${\mu}{\textrm}{m}$) with prokaryotic structure and organization.

• KSBB Journal
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• 제23권5호
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• pp.398-402
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• 2008

Cyanobacteria havebeen identified as one of the most promising group producing novel biochemically active natural products. Cyanobacteria are a very old group of prokaryotic organisms that produce very diverse secondary metabolites, especially non-ribosomal peptide and polyketide structures. Though many useful natural products have been identified in cyanobacterial biomass, cyanobacteria produce also extracellular proteins related with NRPS/PKS. Detection of unknown secondary metabolites in medium was carried in the present study by a screening of 98 cyanobacterial strains. A degenerated PCR technique as molecular approaches was used for general screening of NRPS/PKS gene in cyanobacteria. A putative PKS gene was detected by DKF/DKR primer in 38 strains (38.8%) and PCR amplicons resulted from a presence of NRPS gene were showed by MTF2/MTR2 primer in 30 strains (30.6%) and by A3/A7 primer in 26 strains (26.5%). HPLC analysis for a detection of natural products was performed in extracts from medium in which cyanobacteria containing putative PKS or NRPS were cultivated. CBT57, CBT62, CBT590 and CBT632 strains were screened for a production of extracellular natural products. 5 pure substances were detected from medium of these cyanobacteria.

• Journal of Microbiology and Biotechnology
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• 제20권6호
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• pp.974-977
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• 2010

Cyclophilins contain the conserved activity of cis-trans peptidyl-prolyl isomerase, which is implicated in protein folding, and function as molecular chaperones. When the yeast cyclophilin A gene (cpr1) was subcloned into the prokaryotic expression vector pKM260, it was found that the expression of Cpr1 drastically increased the cell viability of E. coli BL21 when under abiotic stress conditions, as in the presence of cadmium, copper, hydrogen peroxide, heat, and SDS. Therefore, this study illustrates the importance of Cpr1 as a molecular chaperone that can improve the cellular stress responses when E. coli cells are exposed to adverse conditions, while also demonstrating its potential to increase the stability of E. coli strains utilized for the production of recombinant proteins.