• Proceedings of the KSAR Conference
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.50-50
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• 2003

모유에 존재하는 유지방구의 막을 구성하는 주된 당단백질인 하나인 lactadherin(과거에는 BA46로 일컬어짐)은 rotavirus에 의한 감염증상을 예방하는 것으로 보고되고 있다. 본 연구에서는 retrovirus vector system을 이용하여 Chinese Hamster Ovary (CHO) 세포에 tetracycline에 의해 발현이 제어되는 promoter 하의 lactadherin 유전자를 전이시킨 후lactadherin이 tetracycline에 의해 발현이 유도되는지의 여부를 실험하였다. 먼저 기초 실험으로 E. coli LacZ유전자를 이용하여 tetracycline에 의한 유도 여부를 조사하였다. Tet-On과 RevTRE-LacZ retrovirus를 co-infection시킨 NIH3T3 세포는 doxycycline (tetracycline 유도체)의 투여량에 비례하여 E. coli LacZ 유전자의 발현 정도가 증가하는 양상을 나타내었는데, 최대의 발현에 대한 doxycycline 농도는 1 $\mu\textrm{g}$/$m\ell$ 이상으로 나타났다. 이 예비실험의 결과를 바탕으로 Tet-On과 RevTRE-Ltd retrovirus vector를 이용하여 사람의 lactadherin 유전자의 유도적 발현을 검정하였는데, CHO 세포에서 lactadherin 유전자의 유도적 발현을 RT-PCR 기법을 이용하여 확인하였다. 표적세포 내에서 외부에서 도입된 유전자가 지속적으로 발현될 경우 심각한 생리적 부작용을 야기시킨다는 사실을 감안할 때, 본 실험의 결과는 유전자 치료와 형질전환동물의 생산에 크게 도움이 될 것으로 예상된다.

• Korean Journal of Microbiology
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• 제29권1호
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• pp.1-7
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• 1991

Human immunodeficiency virus type 1 (HIV-1) causes a deadly infectious disease, Acquired Immunodeficiency Syndrome (ADIS). As a first step to develop a reliable and fast diagnostic procedure for HIV-1 infection, we cloned various immunodominant epitopes of HIV-1 in bacterial expression vectors containing tac or trp promoter. While the protein level of direct expression of gp160 was low, trp E fused gp120, gp41 and p17-p24 were produced at high levels (15-30% of total bacterial proteins) in E. coli. Since gp120 and gp41 contain relatively conserved regions which can react with antibodies in the plasma from most of HIV-1 infected individuals, these expression clones were used for large preparations of HIV-1 antigens.

• Korean Journal of Microbiology
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• 제25권1호
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• pp.52-56
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• 1987

The evidences that Lam B protein of E. coli is used as a receptor for infections of bacteriophage N4 as well as bacteriophage lambda were obtained from the following experimental results. First, all of the isolated lambda resistant dlones possessing foreign DNA fragments in the plasmids were also resistant to bacteriophage N4, but not to bacteriophage $\phi$ 80, T4 and T7. Second, when the plasmid DNA was treated with various restriction enzymes and ligated to delete the total or a portion of the foreign DNA fragments, the deleted plasmids lost the resistant activities to lambda and N4, simultaneously. Third, after amplification of Lam B protein about 200 times by inducing the protein using maltose as a sole carbon source, the host E. coli became sensitive to both lambda and N4.

• Pediatric Infection and Vaccine
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• 제28권3호
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• pp.168-172
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• 2021

Retroperitoneal infections are rarer than intra-abdominal infections in children; however, they can lead to significant morbidity in cases of missed diagnosis. We present a case of retroperitoneal abscess caused by extended-spectrum β-lactamase-producing Escherichia coli in a 10-year-old female adolescent who failed to show clinical improvement following empirical antibiotic therapy.

• Korean Journal of Animal Reproduction
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• 제19권1호
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• pp.35-41
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• 1995

In this study, we have tested whether the retrovirus vector system is applicable in transgenic cattle production. To overcome low infectivity of currently available retrovirus vector system we have directly microinjected retrovirus-producing cells into the perivitelline space of the day 1.5 embryos. The virus-producing cell line was designed to release replication-defective retrovirus encapsidated with Gibbon ape leukemia virus (GaLV) envelope protein. E. coli LacZ gene was used as a marker gene to facilitate evaluation of the transgene expression and X-gal staining at morula or blastocyst stage resulted in expression of E. coli LacZ gene The results in these experiments were summarized as follows : 1. The lowest concentration of polybrene necessary for efficient virus infection was Sf' g/ml. 2. Development rate from day 1.5 embryos microinjected with virus-producing cells to the morulae /blastocysts was 29%. 3. 21% of the morulae /blastocysts were LacZ+. 4. There was no evidence that the retrovirus-producing cells used in this study produced replication-competent retrovirus.

• Journal of Microbiology and Biotechnology
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• 제9권6호
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• pp.826-831
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• 1999

${\beta}-Glucan$, one of the major cell wall components of Saccharomyces cerevisiae, is known to enhance the immune function, especially by activating macrophages. Accordingly, in an effort to develop a safe and efficient immune stimulatory agent, we prepared crude ${\beta}-glucan$ (glucan-p1) and partially purified ${\beta}-glucan$ that was free of mannoproteins (glucan-p2), and evaluated their effect on both the macrophage function and resistance to E. coli-induced peritonitis. To investigate the function of the macrophages, phagocytosis, $TNF-{\alpha}$ secretion, oxygen burst, and the expression of cytokine genes such as $IFN-{\gamma}$ and IL-12 were analyzed. Glucan-p2 markedly stimulated the macrophages with all these parameters. Glucan-p1, however, did not stimulate phagocytosis, yet it induced $TNF-{\alpha}$ secretion, oxygen burst, and the expression of $IFN-{\gamma}$ and IL-12, although less efficiently than glucan-p2. Finally, to test the in vivo protective effect of {\beta}-glucan against infection, the survival of mice from E. coli-induced peritonitis was investigated. After 24 h of the peritoneal challenge of E. coli, all of the mice treated with glucan-p2 survived whereas none survived in the control group. Glucan-p1 showed only a marginal effect in protecting the mice. These results suggest that mannoprotein-free gluean-p2, but not gluean-p1, can serve as an effective immune-stimulating agent.

• Childhood Kidney Diseases
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• 제10권2호
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• pp.182-191
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• 2006

Purpose : We studied the degree of changes in antibiotic sensitivity toward causative organisms, prevalence and clinical manifestations of extended-spectrum $\beta$-lactamase(ESBL)-producers of urinary tract infection(UTI) for a period of three years. This serves to provide useful information in selecting adequate drugs for the treatment of UTI. Methods : We recruited 137 patients who grew more than $10^5$ CFU/mL in their urine culture among 250 patients who visited and were admitted to Handong University's Sunlin Hospital for UTI treatment from January 2003 to December 2005. We retrospectively analyzed the data from the medical records. Results : The common pathogenic organisms were Escherichia coli(65.0%), Klepsiella pneumoniae(14.0%), Enterococcus faecalis(5.8%) and Proteus vulgaris(2.9%) in consecutive order. The prevalence of ESBL-producers among isolated E. coli and K. pneumoniae was 4.5%(4 cases) and 14.3%(2 cases), respectively. The antibiotic sensitivity rates of E. coli were relatively high to amikacin(100%), imipenem(100%), ceftriaxone(95.5%) and tobramycin(91.4%) while relatively low to TMP/SMZ(55.4%), ampicillin/sulbactam(29.4%) and ampicillin(24.2%). Conclusion : The use of ampicillin, ampicillin/sulbactam and TMP/SMZ, which have been the first choices in the treatment of UTI, should be reconsidered due to the low sensitivity rates towards these antibiotics. Due to the high incidence and antibiotic tolerance of ESBL that might have risen from the development of new antibiotics and increased antibiotic use, it is necessary to consider changing the standard antibiotics that have been used in the treatment of UTI.

• Journal of Life Science
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• 제9권5호
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• pp.556-563
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• 1999

Presence of antibody to the capsid protein p24 is the main diagnostic criterion, since this reflects reliable antibody response to HIV infection. However, it takes about 6-8 weeks for antibody production after infection and people who are infected but antibodies are not produced yet are classified as seronegative. Therefore, there is a strong need for an improved diagnostic method for better health security. As a first step for developing such an improved diagnostic system, gag protein of human immunodificiency virus type 1 was expressed in E. coli DH5$\alpha$. The gag fragment of HIV-1 (including a portion of p17 and whole p24) was amplified by polymerase chain reaction (PCR) and BamH I/EcoR I sites were created during PCR. The amplified DNA fragment was cleaved with BamH I/EcoR I and was subcloned into the GEX-2T vector which had been digested with BamHI/EcoRI, resulting gene fusion with gst gene of pGEX-2T. The recombinant DNA was transferred into E. coli DH5$\alpha$. The transformed bacteria were grown at 37$^{\circ}C$ for 3h and protein expression was induced with 0.1mM IPTG at $25^{\circ}C$ for 3h. Recombinant gag protein or GST-gag fusion protein was purified with glutathione-sepharose 4B bead and migrated as a single band when analyzed by 10% polyacrylamide gel. These proteins were confirmed by immunoblotting with anti-GST goat sera or Korean AIDS patients sera. The results of this study establish the expression and single step pulification of HIV-1 gag protein which can specifically bind with Korean AIDS patients sera.

• Korean Journal of Pharmacognosy
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• 제50권3호
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• pp.191-197
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• 2019

Sepsis, an infectious disease, is a life-threatening condition that arises when the response to infection causes injury to tissues and organs. The purpose of this study was to demonstrate whether ABHC-1 and ABHC-2, two functional extracts from herbal complex, have an anti-bacterial effect against Escherchia coli in vivo, in vitro experimental model. ABHC-1 and ABHC-2 showed the antibacterial activity against the bacteria by paper disc method. The minimum inhibitory concentration (MIC) was measured using alamar blue reagent. The MIC was shown at $60{\mu}g/ml$ from ABHC-1 and $500{\mu}g/ml$ from ABHC-2 against E. coli. We next examined the effect of ABHCs on the production of inflammatory cytokine, such as tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), which is related to the induction of inflammation, in RAW 264.7 cell. ABHC-1 and ABHC-2 increased $TNF-{\alpha}$ production of RAW 264.7 cell in a dose-dependent manner while two extract decreased $TNF-{\alpha}$ production in lipopolysaccharide (LPS)-stimulated RAW 264.7 cell in a dose-dependent manner. At a dose of $1{\times}10^8$ E. coli. i.p., non-treated mice were succumbed, while most of mice treated with ABHC-1 were survived. Therefore, our results suggest that ABHC-1 has anti-bacterial activity and can be a novel therapeutic agent against infectious diseases.

• Korean Journal of Veterinary Research
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• 제62권1호
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• pp.4.1-4.5
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• 2022

This paper describes a fatal case of pneumonia in a 14-day-old dog caused by extraintestinal pathogenic Escherichia coli (ExPEC). The necropsy showed that almost all of left lobes of the lungs had dark-red consolidation. A histopathology examination revealed moderate acute fibrino-hemorrhagic necrotizing pneumonia with intralesional bacterial colonies. Non-suppurative epicarditis, congestion in the liver, and necrosis in the white pulp of the spleen also were found. E. coli with cytotoxic necrotizing factor 1 and α-hemolysin was isolated from the lung. This case was confirmed to have fatal pneumonia caused by ExPEC that led to a systemic infection.