• The Journal of Natural Sciences
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• v.16 no.1
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• pp.1-13
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• 2005

The angiotensin I-converting enzyme (ACE) inhibitor has anti-hypertensive effects and has long been used as prevention or remedy of hypertension. This study were carried out to produce and purify a new ACE inhibitor from recombinant E. coli and further elucidate its structure-function relationship. Recombinant pGEX-4T-3 containing ACE inhibitory peptide gene of Saccharomyces cerevisiae was transformed into E. coli BL21(DE3). Glutathione-S transferase (GST) fusion protein from E. Coli BL21(DE3) harboring the recombination pGEX-4T-3 was obtained and the ACE inhibitory peptide was purified with Sephadex G-25 column chromatography. The purified ACE inhibitory peptide was a novel decapeptide with sequence Tyr-Asp-Gly-Gly-Val-Phe -Arg-Val-Tyr-Thr which shows very low similarity to the other ACE inhibitory peptide sequence. The purified ACE inhibitor competitively inhibited ACE.

• 한국생물공학회:학술대회논문집
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• 2002.04a
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• pp.534-537
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• 2002

An agarase gene from Bacillus cereus ASK202 was expressed in high levels by E. coli BL21(DE3)/pEBA1 using pET28a(+) vector system with the inducible T7 promoter in the presence of isopropyl- ${\beta}$ -thiogalactopyranoside. The open reading frame encodes 761 amino acid residues with a calculated molecular weight of 83,300 daltons and a potential signal peptide about 36 amino acid residues at the N-terminus. E. coli BL21(DE3)/pEBA1 produce 1280 unit/ ${\ell}$ of agarase. The optimum physical condition for the agarase activity was pH 5.6, and $40^{\circ}C$, respectively. The agarase activity was stable up pH $4.0{\sim}9.0$ and $4{\sim}40^{\circ}C$. The km and maximum rate of metabolism for agar were 0.068mg/$m{\ell}$ and 0.094mg/$m{\ell}{\cdot}min$, respectively.

• Journal of Life Science
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• v.14 no.2
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• pp.315-319
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• 2004

For antibody development of human serine palmitoyltransferase (SPT, EC 2.3.1.50), SPTLC1 and SPTLC2 genes were subcloned in pRset vector and expressed in E. coli BL21 (DE3)pLys cells. Eucaryotic SPT is a membrane-bound heterodimer enzyme, while all other members are soluble homodimer enzymes. cDNA library were obtained from total RNA from human embryo kidney cell line, HEK293, using RT-PCR and PCR with specific primers was carried out for preparing SPTLC1 and SPTLC2 genes. pRset vector which can express hexahistidine-tag fusion protein was used and the DNA sequences of pRsetB/SPTLC1 and pRsetA/SPTLC2 were confirmed. Recombinant BL21 cells with SPTLC subunits were selected with LB plate containing ampicillin and chroramphenicol. SPTLC1 and SPTLC2 proteins were induced with 1 mM IPTG and seperated on 10% SDS-PAGE gel. Expressed proteins were confirmed by western blotting with His-tag antibody.

• Journal of Acupuncture Research
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• v.21 no.3
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• pp.169-178
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• 2004

Objective : This study was designed to investigate the effects of acupuncture, moxibustion and electro-acupuncture at LI4 and LR3, BL18 and BL25 on serum gastin level in rats. Methods : LI4 and LR3 which belong to Large Intestine Meridian and liver Meridian are acupuncture points frequently used for healing gastrointestinal disease in oriental medicine. BL18 and BL25 which belong to Bladder Meridian are acupuncture points used for healing hepatic disease and intestinal disease in oriental medicine. Serum gastrin level by radioimmnunoassay was measured at 3 days after acupuncture, moxibustion and electro-acupuncture of those acupoints. Results & Conclusion : Moxibustion applied to the BL18 acupoint, BL25 acupoint decreased gastrin level of serum, but moxibustion applied to the BL18+BL25 acupoints did not produced significant effect. Acupuncture applied to the LI4 acupoint, LR4 acupoint did not produced significant effect, but acupuncture applied to the LI4+LR4 acupoints increased gastrin level of serum, Electro-acupuncture applied to the LI4 acupoint, LR4 acupoint increased gastrin level of serum, and electro-acupuncture applied to the LI4+LR4 acupoints also increased gastrin level of serum.

• Biomolecules & Therapeutics
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• v.21 no.1
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• pp.66-71
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• 2013

Broussonetia papyrifera and Lonicera japonica have long been used in the treatment of inflammatory disorders in Chinese medicine, especially respiratory inflammation. Previously, a new phytoformula (BL) containing B. papyrifera and L. japonica was found to exert strong anti-inflammatory activity against several animal models of inflammation, especially against an animal model of acute bronchitis. In the present investigation, the effects of BL on animal models of septic inflammation and chronic bronchitis are examined. Against lipopolysaccharide (LPS)-induced septic inflammation in mice, BL (200-400 mg/kg) reduced the induction of some important proinflammatory cytokines. At 1 h after LPS treatment, BL was found to considerably inhibit TNF-${\alpha}$ production when measured by cytokine array. At 3 h after LPS treatment, BL inhibited the induction of several proinflammatory cytokines, including IFN-${\gamma}$ and IL-$1{\beta}$, although dexamethasone, which was used as a reference, showed a higher inhibitory action on these biomarkers. Against chronic bronchitis induced by LPS/elastase instillation in rats for 4 weeks, BL (200-400 mg/kg/day) significantly inhibited cell recruitment in bronchoalveolar lavage fluid. Furthermore, BL considerably reduced lung injury, as revealed by histological observation. Taken together, these results indicate that BL may have a potential to treat systemic septic inflammation as well as chronic bronchitis.

• Korean Journal of Acupuncture
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• v.18 no.1
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• pp.117-141
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• 2001

From documentary comparative study on the chief virtues of the Back-Su(兪) points and the Front-Mo(募) points, the following results are obtained : 1. Contrast the Back-Su(兪) points with the Front-Mo(募) points, the needle-steadying depth of the Front-Mo(募) points is deeper than that of the Back-Su(兪) points. and Moxibustion dosage of the Front-Mo(募) points is more than that of the Back-Su(兪) points. 2. In the case of Kimun(LR14) and Kansu(BL18), Kergwol(CV12) and Shimsu(BL15), Chungwan(CV12) and Wisu(BL21), Kyungmun(GB25) and Shinsu(BL23), the Back-Su(兪) points can be chiefly used for the treatment of acute disease, external disease, exess disease and Jang(臟)-disease. contrary the Front-Mo(募) points can be chiefly used for the treatment of chronic disease, internal disease, deficiency disease and Bu(腑)-disease. 3. In the case of Kwangwon(CV4) and Sojangsu(BL27), Serkmoon(CV5) and Samchosu(BL22), Changmun(LR13) and Bisu(BL20), Chungbu(LV1) and Pesu(BL20), Chungguk(CV3) and Banggwangsu(BL28), the Front-Mo(募) points can be used for the treatment of unbalanced visceral funtion. contrary the Back-Su(兪) points can be used for not only the treatment of unbalanced visceral funtion but also the treatment of attached organs. 4. The Back-Su(兪) points can be used for the treatment of acute disease, external disease, exess disease, Jang(臟)-disease and attached organs-disease. contrary the Front-Mo(募) points can be chiefly used for the treatment of chronic disease, internal disease, deficiency disease, and Bu(腑)-disease.

• Microbiology and Biotechnology Letters
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• v.32 no.2
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• pp.135-141
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• 2004

Human granulocyte colony-stimulating factor (hG-CSF) is a hematopoiesis agent that principally affects the differentiation of neutrophils in the bone marrow. At present, recombinant hG-CSF is used successfully in the treatment of chemotheraphy-induced neutropenia and its indication has been expanded to bone marrow transplantation and aplastic anemia. In this study, we have constructed rhG-CSF secretion plasmid pYRC1 in which OmpA signal sequence/hG-CSF gene was expressed under the control of the T7 promoter. rhG-CSF produced in E. coli BL21 (pYRC1) grown at $37{\circ}C$ was found in aggregates. However, 15% of the periplasmic protein was soluble rhG-CSF when the E. coli BL21 (pYRC1) was cultured at $25^{\circ}C$ for 7 h in the modified MBL medium containing 10 g/$\ell$ glucose with 10 $\mu$M IPTG induction. The production of soluble rhG-CSF in E. coli BL21 (pYRC1) using fed batch culture was also studied. In the fed batch culture system, the final yield of rhG-CSF produced from E. coli BL21 (pYRC1) was increased from 4.4 mg/$\ell$to 24 mg/$\ell$by controlling the specific growth rate from $0.43 h^{-1}$ to $0.14 h^{-1}$, and optimizing the time of induction.

• KSBB Journal
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• v.13 no.6
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• pp.662-668
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• 1998

Viruses belonging to the Hantavirus genus cause two acute severe illness in humans, i.e., Haemorrhagic Fever with Renal Syndrome (HFRS) and Hantavirus Pulmonary Syndrome(HPS). Among them, Hantaan virus is one of the most important viruses causing HFRS. Recombinant expression vectors, pKK-NP and pET-NP, with Hantaan viral nucleocapsid gene were constructed, and used to transform Eschericia coli BL21(DE3). Stability of the vectors in the host strain, and effects of some environmental conditions on the expression of nucleocapsid gene were studied. Expression vector, pKK-NP, was very unstable, and the expression level of nucleocapsid gene was very low compared to that of pET-NP. BL21(pET-NP) produced about 100 mg of N protein per liter of culture broth. Induction time did not show any significant difference on the expression level of nucleocapsid gen and cell growth. BL21(pET-NP) culture at 35$^{\circ}C$ showed a little higher expression level than at 30$^{\circ}C$ during growth phase, but reached to the same level at stationary phase. Total expression level was proportional to supplemented glucose concentration of media up to 0.5% along with cell growth, but expression level per unit cell mass was inversely proportional to glucose concentration and maximal when glucose was not supplemented at all.

• Journal of Veterinary Clinics
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• v.20 no.1
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• pp.49-51
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• 2003

This study was performed to evaluate the effect of acupuncture on gastric motility in dogs with radiography and ultrasonography. All experimental dogs were fed with the soup with barium (3% W/V), and thereafter gastric emptying time(GET) was measured with radiography and ultrasonography with Bolondi's method. The acupoints were CV-12, PC-6, ST-36 and BL-21 in this experiment. On these acupoints, electroacupunctral stimulation(15 MHz, continous wave) was applied for 20 minutes. The results were as follows: In the control group, normal GET was 70-85 minutes with ultrasonography, and 110-130 minutes with radiography. In acupuncture stimulation groups, there was no effect of gastric motility in CV-12, PC-6, and 57-36, whereas there was significant stimulating effect of gastric movement in BL-21 with ultrasonography (P < 0.05) and radiography (P < 0.01). Above these results, it is concluded that acupunctural stimulation of BL-21 was more effective in gastric motility compared to those of CV-l2, PC-6 and ST-36.

• Journal of Integrative Natural Science
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• v.1 no.1
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• pp.24-31
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• 2008

Brassinosteroids (BRs) are a special class of plant steroid hormones that are essential for normal growth and development. Part of confusion is whether BRs are unique to plants, because they have overlapping physiological roles with other better-studied hormones and with physiological responses caused by light. In systems designed to assay for cytokinins, the effects of BRs vary. We measured hypocotyl length for testing the ability of brassinolide (BL) to rescue double mutant between det2 and the photoreceptor null mutant phytochrome B (phyB). PHYB involved in controlling hypocotyl elongation in increased concentration of BL whereas phyBdet2 double mutant just partially rescue to phyB in white and red light indicated the involvement of BRs in PHYB regulated cell elongation. BRs regulated hypocotyl growth was delayed by BAP, a cytokinin treatment but inhibitory effects of BAPs on hypocotyl growth was slightly recovered by BL. The result indicated that the mode of action of BR and cytokinin is independent or sequential in the downstream light-regulated response control on hypocotyl elongation and also light modulated the action of BR and cytokinin in some extent.