• Journal of Microbiology
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• v.33 no.3
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• pp.191-195
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• 1995

IciA protein has been shown as an inhibitor for the initiation of E. coli chromosomal DNA replication at oriC. IciA protein binds the AT-rich region in oriC and then blocks the initiation of chromosomal DNA replication. Two binding sites for IciA protein were identified in dnaA gene, encoding the initiator for the E. coli chromosomal replication, promoter region by gel-shift assay and DNase I footprinting, One, named as IciA site I, is located upstream of the dnaA promoter 1P. The other, named as IciA site II, is located downstream of the dnaA promoter 2P. The sequence comparison of the regions protected from the DNase I cleavage did not result in a clear consensus sequence for the binding of IciA protein, suggesting that IciA protein may be a member of multimeric complex dsDNA binding proteins. This study provided information about the binding mode of IciA protein. Even though the IciA site II and IciA binding site in oriC seem to be composed of two IciA binding units, one binding unit is likely enough to cause the binding of IciA protein to the IciA site I. The binding of IciA protein to the dna4 promoter implies that IciA protein may involve not only the control of the initiation of chromosomal DNA replication but also the control of the dna4 gene expression.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.27 no.2
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• pp.103-110
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• 2001

This study was performed to observe the effect of ultrasound(1.0MHz, $0.75W/cm^2\;and\;1.0W/cm^2$) irradiation on cultured MC3T3-E1 cell, osteoblastic like cell with respect to the proliferation, protein synthesis, and alkaline phosphatase activity of the cells. The results were as follows: 1. The proliferation of MC3T3-E1 cells was increased on ultrasound irradiated group compared with control group. 2. The protein synthesis was not apparently increased on ultrasound irradiated group compared with control group. 3. The alkaline phosphatase activity level was not apparently increased on ultrasound irradiated group compared with control group. From the above results and other literatures, we could suggest that the ultrasound with the appropriate intensity and frequency may have important roles in stimulation of cell proliferation. Therefore the ultrasound may be used in the acceleration of the bone regeneration and bone fracture healing.

• The Plant Pathology Journal
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• v.21 no.4
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• pp.334-342
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• 2005

The prototype Chlorella virus PBCV-l encodes 11 tRNA genes and over 350 protein-encoding genes in its 330 kbp genome. Initial attempts to overexpress the recombinant A189/192R protein, a putative virus attachment protein, in E. coli strain BL21(DE3) SI were unsuccessful, and multiple protein bands were detected on Western blots. However, the full-length A189/192R recombinant protein or fragments derived from it were detected when they were expressed in E. coli BL21 CodonPlus (DE3) RIL, which contains extra tRNAs. Codon usage analysis of the a189/192r gene showed highly biased usage of the AGA and AVA codons compared to genes encoded by E. coli and Chlorella. In addition, there were biases of XXA/U($56\%$) and XXG/ C($44\%$) in the codons recognized by the viral tRNAs, which correspond to the codon usage bias in the PBCV-1 genome of XXA/U ($63\%$) over those ending in XXC/G ($37\%$). Analysis of the codon usage in the major capsid protein and DNA polymerase showed preferential usage of codons that can be recognized by the viral tRNAs. The Asn (AAC) and Lys (AAG) codons whose corresponding tRNA genes are duplicated in the tRNA gene cluster were the most abundant (i.e., preferred) codons in these two proteins. The tRNA genes encoded in the PBCV-l genome seem to play a very important role during the synthesis of viral proteins through supplementing the tRNAs that are frequently used in viral proteins, but are rare in the host cells. In addition, these tRNAs would help the virus to adapt to a wide range of hosts by providing tRNAs that are rare in the host cells.

• Korean Journal of Environmental Biology
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• v.23 no.3 s.59
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• pp.275-280
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• 2005

Development of the fourth-instar larvae of Chironomus riparius has a sensitive to ecdysteroid hormones. The 2D/E gel analysis for polypeptide expression reflecting early-ecdysterone inducible gene has conducted the emerged female from larval phase exposure to bisphenol A (BPA). In the 2D/E gel 1108 protein spots were identified. The visualized protein spots allowed extraction of 17 protein spots differed more than 3 fold in BPA treated animals, which was approximately $1.6\%$ of the total protein spots. However, polypeptide expression reflecting early-ecdysterone inducible gene didn't change after treatments. In addition, detection for the damages or changes in mitogenome level was observed. The conserved cytochrome oxidase I in DNA level affected exposure to BPA $(1{\mu}gL^{-1})$ in this preliminary study.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.809-819
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• 1996

This study was conducted to identify the effects of caffeine or combinations of caffeine and iron or vitamin E on the lipid and protein components in the MDBK(Mardin-Darby Bovine Kidney) cells. For the In vitro test, MDBK cells in ${\alpha}$-MEM(Minimum Essential Medium) were divided into 4 treatment groups according to drug types and dosages as follows; the control(group A), group B was treated with 0.3mM caffeine, group C was treated with 0.3mM caffeine and 0.3mM ferric chloride, group D was treated with 0.3mM caffeine and 0.3mM vitamin E. Those groups were further divided into 5 subgroups according to the time lapsed(control, 4hrs, 8hrs, 24hrs and 48hrs lapsed group). The concentrations of the carbonyl group and malondialdehyde(MDA) and the patterns of the SDS-PAGE(Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) and fatty acid compositions were analyzed to determine the oxidative damages and metabolic changes on the lipid and protein components in the MDBK cells. The results obtained from this study were summarized as follows; 1. The concentrations of carbonyl group and malondialdehyde in MDBK cells of group C were significantly higher(p<0.01) in comparison to the control, and increased according to the time lapsed. But the results of groups B and D were little different in comparison to the group C. 2. As the analytical results of fatty acid compositions in MDBK cells, the proportions of palmitoleic acid and linoleic acid in groups B, C and D were lower in comparison to the control, while the proportion of arachidonic acid in groups B, C and D were significantly higher(p<0.01) in comparison to the control. 3. In order to determine the oxidative damages to the protein in MDBK cells, the patterns of the SDS-PAGE were examined and the patterns of SDS-PAGE in groups C and D were significantly different between 43kd and 200kd of molecular weight.

• Biomolecules & Therapeutics
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• v.4 no.1
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• pp.97-102
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• 1996

To investigate whether human $\beta$$_2$-adrenergic receptor devoid of the C-terminal two transmembrane helices retain its ligand binding activity and specificity, 5'780-bp DNA fragment of the receptor gene which encodes amino acid 1-260 of human $\beta$$_2$-adrenergic receptor was subcloned into the bacterial fusion protein expression vector and expressed as a form of glutathione-S-transferase (GST) fusion protein in E. coli DH5$\alpha$. The receptor fusion protein was expressed as a membrane bound form which was verified by SDS-PAGE and Western blot. The fusion protein expressed in this study specifically bound $\beta$-adrenergic receptor ligand [$^3$H] Dihydroalprenolol. In saturation ligand binding assay, the $K_{d}$ value was 7.6 nM which was similar to that of intact $\beta$$_2$-adrenergic receptor in normal animal tissue ( $K_{d}$=1~2 nM) and the $B_{max}$ value was 266 fmol/mg membrane protein. In competition binding assay, the order of binding affinity of various adrenergic receptor agonists to the fusion protein was isoproterenol》epinephrine norepinephrine, which was similar to that of intact receptor in normal animal tissue. These results suggest that N-terminal five transmembrane helices of the $\beta$$_2$-adrenergic receptor be sufficient to determine the ligand binding activity and specificity, irrespective of the presence or absence of the C-terminal two transmembrane helices.s.s.s.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.30 no.4 s.48
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• pp.479-483
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• 2004

An attempt was made to develop a proteinchip for screening of MITF (microphthalmia transcription factor) inhibitor. Binding of MITF to E-box causes transcription of several pigmenting genes including tyrosinase gene. We investigated binding of MITF and its DNA binding site (E-box) using a protein-DNA chip with various detection methods including flurorescence (Cyt3). SPR (surface plasmon resonance) and SPRi (surface plasmon resonance imaging). A fusion protein (MITF-Maltose Binding Protein) was attached on the glass plate by chemical modification. An inhibitory synthetic DNA oligomer, artificially designed based on the E-box sequence, inhibited the binding of MITF and E-box. These results showed the potentials of flurorescence-based MITF protein chip as a microarray for high throughput screening (HTS) system of depigmenting agents.

• Biomolecules & Therapeutics
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• v.11 no.3
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• pp.157-162
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• 2003

The generation of secretory IgA antibodies(Abs) for specific immune protection of mucosal surfaces depends on stimulation of the mucosal immune system, but this is not effectively achieved by parenteral or even oral administration of most soluble antigens. Thus, to produce a possible vaccine antigen against urinary tract infections, the uropathogenic E. coli (UPEC) adhesin was genetically coupled to the ctxa2b gene and cloned into a pMAL-p2E expression vector. The chimeric construction of pMALfimHIctxa2b was then transformed into E. coli K-12 TB1 and its nucleotide sequence was verified. The chimeric protein was then purified by applying the affinity chromatography. The purified chimeric protein was confirmed by SDS-PAGE and western blotting using antibodies to the maltose binding protein (MBP) or the cholera toxin subunit B (CTXB), plus the N-terminal amino acid sequence was analyzed. The orderly-assembled chimeric protein was confirmed by a modified $G_{M1}$-ganglioside ELISA using antibodies to adhesin. The results indicate that the purified chimeric protein was an Adhesin/CTXA2B protein containing UPEC adhesin and the $G_{M1}$-ganglioside binding activity of CTXB. This study also demonstrate that peroral administration of this chimeric immunogen in mice elicited high level of secretory IgA and serum IgG Abs to the UPEC adhesin. The results suggest that the genetically linked CTXA2B acts as a useful mucosal adjuvant, and that the adhesin/CTXA2B chimeric protein might be a potential antigen for oral immunization against UPEC.

• The Journal of Korean Medicine
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• v.43 no.3
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• pp.106-121
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• 2022

Objectives: The purpose of this study was to investigate the anti-inflammatory effect of Cornus Officinalis fruit extract(CE) and Cornus Officinalis Fruit Cheonghyeol Plus(CCP) in Human Umbilical Vein Endothelial Cell. Methods: We measured cell viability of CE, CCP and treated HUVEC with TNF-α. We measured the mRNA expression levels of KLF2, eNOS, MCP-1, ICAM-1, VCAM-1, the protein expression levels of KLF2, eNOS, MCP-1, ICAM-1, VCAM-1, and the protein phosphorylation level of ERK, JNK, p38 and the biomarker expression levels of MCP-1, ICAM-1, VCAM-1. Results: 1.CE incresed the mRNA, protein expression levels of KLF2, eNOS at concentrations of 100㎍/㎖ compared to the control group. CE decresed the mRNA, protein and biomarker expression levels of MCP-1,ICAM-1,VCAM-1 at concentrations of 100㎍/㎖ compared to the control group. CE decresed the protein phosphorylation level of p38 at concentrations of 100㎍/㎖ compared to the control group. 2. CCP incresed the mRNA, protein expression levels of KLF2, eNOS at concentrations of 100㎍/㎖ or more compared to the control group. CCP decresed the mRNA, protein and biomarker expression levels of MCP-1, ICAM-1, VCAM-1 at concentrations of 100㎍/㎖ or more compared to the control group. CCP decresed the protein phosphorylation level of ERK at concentrations of 100㎍/㎖ or more, p38 at concentrations of 200㎍/㎖ or more, and JNK at concentrations of 400㎍/㎖ compared to the control group. Conclusions: These results present that CE and CCP has anti-inflammatory effect in HUVEC. So, it could help treat or prevent inflammation in vein caused by dyslipidemia and contribute prevention of cardiovascular and cerebrovascular cerebrovascular diseases.

• Journal of Microbiology and Biotechnology
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• v.18 no.7
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• pp.1235-1244
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• 2008

Indole-3-acetic acid (IAA) is produced commonly by plants and many bacteria, however, little is known about the genetic basis involving the key enzymes of IAA biosynthetic pathways from Bacillus spp. IAA intermediates from the Gram-positive spore-forming bacterium Paenibacillus polymyxa E681 were investigated, which showed the existence of only an indole-3-pyruvic acid (IPA) pathway for IAA biosynthesis from the bacterium. Four open reading frames (ORFs) encoding indole-3-pyruvate decarboxylase-like proteins and putative indole-3-pyruvate decarboxylase (IPDC), a key enzyme in the IPA synthetic pathway, were found on the genome sequence database of P. polymyxa and cloned in Escherichia coli DH5$\alpha$. One of the ORFs, PP2_01257, was assigned as probable indole-3-pyruvate decarboxylase. The ORF consisted of 1,743 nucleotides encoding 581 amino acids with a deduced molecular mass of 63,380 Da. Alignment studies of the deduced amino acid sequence of the ORF with known IPDC sequences revealed conservation of several amino acids in PP2_01257, essential for substrate and cofactor binding. Recombinant protein, gene product of the ORF PP2_01257 from P. polymyxa E681, was expressed in E. coli BL21 (DE3) as a glutathione S-transferase (GST)-fusion protein and purified to homogeneity using affinity chromatography. The molecular mass of the purified enzyme showed about 63 kDa, corresponding closely to the expected molecular mass of IPDC. The indole-3-pyruvate decarboxylase activity of the recombinant protein, detected by HPLC, using IPA substrate in the enzyme reaction confirmed the identity and functionality of the enzyme IPDC from the E681 strain.