• Biomolecules & Therapeutics
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• 제6권3호
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• pp.276-282
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• 1998

Retinoids regulate a wide variety of biological processes such as cellular proliferation and differentiation in many cell types. They have also shown to stimulate replication of several viruses including human cytomegalovirus (CMV). Retinoid signalling pathway involves two distinct subfamilies of nuclear receptors, retinoic acid receptors (RARs) and retinoid X receptors (RXRs) that bind to specific retinoic acid response elements (RAREs) in the promoter regions of retinoid-target genes. Here, we characterized RAREs in the regulatory regions of the CMV and of the hepatitis B vi.us (HBV). The viral RAREs, i.e., CMV-RARE and HBV-RARE, are composed of two consensus RARE half-sites (A/GGGTCA) arranged as a direct repeat separated by 5-bp and 1-bp, respectively. The RAREs were activated by both RAR/RXR heterodimers and RXR homodimers in transient transfection experiments. We also found that COUP-TF$\alpha$ (chicken ovalbumin upstream promoter-transcription factor u) and COUP-TF$\beta$ repressed the retinoid response of the viral elements. Further we demonstrated that previously known retinoid antagonist, SRI 1330, repressed retinoid-induced transactivation of the CMV-RARE. These results implicate Vitamin A, it's nuclear receptors and COUP-TFs as important regulators of the CMV and HBV pathogenesis and the SRl1330 as potential negative modulator of such retinoid-dependent processes.

• 한국정보통신학회:학술대회논문집
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• 한국정보통신학회 2013년도 추계학술대회
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• pp.946-948
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• 2013

베큘로바이러스 시스템이 제조되었는데 이것은 polyhedron promoter, vesicular stomatitis virus G (VSVG), polyA, cytomegalovirus (CMV) promoter, enhanced green fluorescent protein (EGFP), protein transduction domain (PTD) 유전자를 재조합한 것이다. 본 재조합벡터 시스템은 인간 섬유아세포에 적용하여 시험하였고 재조합된 유전자의 전이와 유전자 발현을 대조 벡터시스템과 비교하였다. 본 연구로부터 새롭게 제작된 본 베큘로바이러스 시스템이 유전자의 전이와 유전자 발현 면에서 대조 벡터시스템 보다 고효율을 나타내었다.

• 약학회지
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• 제50권4호
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• pp.268-271
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• 2006

Protein kinase UL97 is an unusual protein kinase that can phosphorylate nucleoside analogs as well as protein/peptide. Previously we found a H2B-derived peptide, KESYSVYVYKV and reported that the P+5 position (K) is important. To further understand the substrate specificity at the P+5 position, we introduced spot assay system and showed that a peptide containing K residue among other amino acids at the P+5 position is the best substrate. Also other residues such as M, I, L, or G are good enough to be substrate of UL97. This result may aid the discovery of a new antiviral inhibitor.

• Journal of Microbiology
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• 제39권3호
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• pp.226-228
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• 2001

Human lymphotropic herpesvirus is known to be a major pathogen associated with various diseases in bone marrow transplantation (BMT) recipients. A multiplex nested-polymerase chain reaction (PCR) method was developed for the simultaneous detection of human lymphotropic herpesviruses, including Ebstein-Barr virus (EBV), cytomegalovirus (CMV), and human herpesvirus 6 variants A and B (HHV6-A, HHV6-B). To demonstrate the usefulness of multiplex PCR for the analysis of clinical samples, peripheral blood mononuclear cells and serum from BMT recipients were analysed. The results skewed that a clear detection could be made between EBV, HCMV and HHV-6. This multiplex PCR assay is an efficient and cost-effective approach to the analysis of large numbers of samples to determine the epidemiological importance of EBV HCMV and HHV-6.

• Annals of Clinical Neurophysiology
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• 제7권1호
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• pp.25-27
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• 2005

The Guillain-Barre syndrome (GBS) is an acute polyradiculoneuropathy marked by flaccid areflexic paralysis. Although the pathogenesis of GBS remains incompletely defined, considered as an autoimmune disease most frequently triggered by an previous infection. Antecedent infections with Campylobacter jejuni, cytomegalovirus, Ebstein-Barr virus, Mycoplasma pneumoniae, Haemophilus influenzae, human immunodeficiency virus, enterovirus, rotavirus are common. But, it is rare that GBS following typhoid fever. We present a case of typical GBS after antecedent Salmonella typhi infection.

• 약학회지
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• 제52권6호
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• pp.466-470
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• 2008

Human cytomegalovirus expresses an unusual protein kinase UL97, a member of ${H_V}{U_L}$ family of protein kinase. UL97 can phosphorylate nucleoside analogs such as ganciclovir as well as protein/peptide. It has previously been reported that UL97 is able to phosphorylate a KESYSVYVYKV peptide and that P+5 position (K) is important. We examined the extent of contribution of other positions (P-4 through P+6) of the peptide to be substrate of UL97 using alanine substituted peptides (Ala scanning) and deleted peptides. The result suggested that the E (P-2) is negative effect and P+5 (K) is still important. The peptide YSVYVYK is the shortest substrate enough to show high activity, which could be a starting point to develop peptidomimetic drug. This study would give important information to deeply understand the substrate specificity of UL97 and develop an antiviral drug using the small peptide identified here.

• 농업과학연구
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• 제48권2호
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• pp.353-365
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• 2021

Previously, we reported that tissue factor (Tf) was included in the list of differentially expressed genes as an upregulated gene in a rejected porcine heart after xenotransplantation into monkey. In this study, we analyzed that expression of Tf in aortic endothelial cells (pAEC) isolated from alpha 1,3-galactosyltransferase knockout pig in response to allogeneic porcine serum and xenogeneic human serum. The consequence was significant upregulation of Tf expression by responding to human serum compared with porcine serum. To analyze the function of Tf gene as a promoter, we constructed reporter vectors for expression of luciferase linked to 1,246 and 787 base pairs of porcine Tf (pTF1246 and pTF787), and 535 base pairs of human TF (hTF535) sequences including putative promoter regions and AP-1 biding site at the 5' end. The reporter vectors were transfected into pAEC including cytomegalovirus enhancer/chicken β-actin (CAG)-luciferase vector as a control. Luciferase assay showed that all of the promoters were insufficient to express luciferase compared with CAG promoter in basic culture conditions. Notably, pTF1246, pTF787, and hTF535 led to a significant increase of luciferase expression in response to human serum compared with porcine serum while no change of CAG. pTF1246 and pTF787 showed higher expression than hTF535. Taken together, our findings suggest that pTF1246 and pTF787 promoters could mediate target gene expression specifically at xenogeneic stress condition.

• Pediatric Infection and Vaccine
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• 제6권2호
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• pp.253-260
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• 1999

목 적 : human CMV가 생성하는 분자량 72,000 dalton의 immediate early protein(p72)과 반응하는 단세포군 항체를 국내에서 분리된 CMV 야생주로 직접 제작하여 진단에 사용해 보기 위한 것이다. 방 법 : 정상 인체 섬유아세포(Foreskin 유래)를 10% 우태아혈청이 함유된 Eagle' s minimal essential medium(MEM)에서 배양하여 사용하였으며 마우스 골수종세포는 P3X63 Ag8.653(ATCC, Mary land USA)을 $5{\times}10^5/ml$ 세포수를 유지하도록 배양하였다. 사용된 항원은 선천성 CMV 감염아의 소변으로부터 검출한 CMV(KJHJ90)를 대량 증식시켜 사용하였다. 생후 6~8주된 Balb/c마우스의 복강내에 CMV 항원을 1주일 간격으로 4회 주사한 후 면역시킨 마우스 비장에서 얻은 임파구와 형질세포종세포로 최적발육기의 P3X63-Ag8.653을 사용하여 융합하였다. 융합세포를 배양한 후 간접면역형광항체법을 이용하여 항체가가 높은 것을 골랐다. 결 과 : 생산된 28종의 단세포군항체 중 그림 1과 같이 LPC12와 LPC23클론이 AD169에 감염된 세포의 핵에 특히 강하게 반응하였다. 정제된 AD169를 SDS-PAGE한 후 Western blotting을 하여 생산된 단세포군항체가 반응하는 항원의 분자량을 확인하였으며 그 중 LPC12와 LPC23클론에서 생성된 단세포군항체는 약 72KDA의 항원과 특이적으로 가장 강하게 반응하였다. 결 론 : 본 연구에서 제작된 LPC12와 LPC23 클론에서 생성된 단세포군 항체를 이용하여 선천성 CMV감염으로 확인된 소아의 소변을 이용하여 CMV AD 169 주 유래 항체(P63-27)과 비교한 DEAFF 검사상 동일한 결과를 확인한 바 LPC12와 LPC 23 클론 항체는 AD 169(P63-27)과 같이 CMV 감염의 조기진단에 사용할 수 있을 것으로 사료된다.

• Reproductive and Developmental Biology
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• 제28권3호
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• pp.191-196
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• 2004

본 실험에서는 외래 유전자의 효율적인 발현을 위하여 GFP 표지유전자를 이용하여 여러 종류의 promoter를 검정하였다. 또한, retrovirus vector에 WPRE 서열을 도입함으로써 GFP 유전자의 발현 증가 여부를 확인하였다. 모든 표적 세포에 있어서 UbC와 β-actin promoter에 비해 RSV와 CMV promoter 통제하의 GFP의 발현이 더 강하게 나타났으며, 특히 CEF 세포에서는 RSV promoter가 가장 우수한 것으로 확인되었다. WPRE의 도입으로 인한 발현율의 증가는 CEF를 제외한 세포주에서 promoter의 종류에 관계없이 확인되었다. 이상의 결과로 각 세포주는 promoter에 따라 발현 양상이 약간의 차이를 보이고 있으나 RSV와 CMV promoter에서 유전자의 발현이 보다 효율적이며, WPRE 서열이 도입된 경우에 HeLa와 PFF 세포에서 발현이 현저히 증가하는 것을 확인할 수 있었다. 이러한 연구 결과는 효율적인 유전자의 발현 체계를 확립하는데 기여함으로써 더 나아가 유전자 치료나 형질전환 동물생산에 적극적으로 활용되어질 수 있을 것이다.

• The Korean Journal of Physiology and Pharmacology
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• 제3권6호
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• pp.597-603
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• 1999

Caldesmon (CaD), one of microfilament-associated proteins, plays a key role in microfilament assembly in mitosis. We have investigated the effects of overexpression of the high molecular weight isoform of CaD (h-CaD) on the physiology of vascular smooth muscle cells (VSMCs). Rat aortic VSMCs were stably transfected with plasmids carrying a full length human h-CaD cDNA under control of cytomegalovirus promoter. The majority of the overexpressed h-CaD appears to be localized predominantly on cytoskeleton structures as determined by detergent lysis. The overexpression of h-CaD, however, does not decrease the level of endogenous low molecular weight isoform of CaD. h-CaD overexpressing VSMCs (h-CaD/VSMCs) show a decreased growth rate than that of vector-only transfected cells when determined by $[^3H]thymidine$ uptake and cell counting after fetal bovine serum (FBS) stimulation. h-CaD/VSMCs were smaller than vector-transfected cells by 18% in cell diameter. These data suggest that overexpression of h-CaD can inhibit the poliferation and the cell volume of VSMCs stimulated by growth factors and that the gene therapy with h-CaD may be helpful to prevent the conditions associated with hypertrophy and/or hyperplasia of VSMCs after arterial injuries.