Activation of the Human Endogenous Retrovirus W Long Terminal Repeat by Herpes Simplex Virus Type 1 Immediate Early Protein 1

장경립,Woo Jung Lee,Hyun Jin Kwun,김희수 한국분자세포생물학회 2003 Molecules and cells Vol.15 No.1

수계 장바이러스의 효과적인 농축과 검출방법의 개발

장경립,정은영 한국생명과학회 2000 생명과학회지 Vol.10 No.4

Enteroviruses in the environment pose a public health risk because they can be transmitted via the fecal-oral route through contaminated water, and low numbers are able to initiate an infection in humans. Because the levels of viruses typically found in environmental water and drinking water are low, they must be concentrated from hundreds to thousands of liters of water. Therefore, the main goal of this study was the development of a rapid, simple and efficient procedure to concentrate, isolate and detect enteroviruses from environmental water samples. Viruses were first concentrated by adsorption to 1 MDS cartridge filter and then eluted with approximately 0.5 liter of 1.5% beef extract/0.05M glycin(pH 9.4). In this study, several procedures to concentrate and purify intact viruses from beef extract obtained from the adsorbent filters were tested. Among them, organic floccuration was the best reliable method for reconcentration. sample volume could be reduced to 200∼400 folds and the efficiency of virus recovery through the procedure was over 72%. Finally, the samples were filtered through a membrane disk filter and then analyzed by either the plaque assay or combined cell culture-polymerase chain reaction.

Repression of p21 Expression by Hepatitis B Virus X Protein via a p53-Independent Pathway

안지영,장경립 한국생명과학회 2000 한국생명과학회 학술발표회 Vol.30 No.-

한국인 간염환자에서 분리한 G형 감염바이러스(HGV)의 외피영역의 유전적 다양성 분석

김종경,장경립 한국생명과학회 1998 생명과학회지 Vol.8 No.4

The genetic of a recently described virus, hepatitis G virus(HGV) was investigated. HGV envelope 1 (E1) nucleotide sequences isolated from six Korean hepatitis b virus-positive patients by using a reverse transcription-poly-merase chain reaction procedure, were analysed and compared to the seven previously reported HGV isolates. Sequence homology among the Korean isolates was 88-97% whereas among the isolates from different geographic areas was 80-92%, indicating geographical divergence of HGV. Nucleotide substitutions spread uniformly throughiut the E1 fragment. Furthermore, compared to the prototype HGV sequence, frameshift mutations were observed in most of the Korean isolating that a different translating initiation site for the polyprotein exists in the Korean type HGV.

대하새우로부터 분리한 WSBV 의 게놈서열 분석

이태호,장경립,전홍기,손상규,허문수,김종경 한국어병학회 1997 한국어병학회지 Vol.10 No.2

새우의 갑각에 흰점을 유발하는 특징을 가진 WSBV는 Baculovirus의 일종으로 여러 종류의 새우에 높은 치사율을 보이는 병원체로서 새우양식에 막대한 피해를 주고 있다. 본 연구에서는 국내에서 양식중인 대하에 질병을 유발한 WSBV의 특성을 알아내고자 치사한 새우로부터 바이러스의 게놈을 클로닝하여 재조합클론(E3)을 분자생물학적으로 분석하였다. E3의 염기서열을 분석한 결과, 이 클론은 AcNPV를 포함한 지금까지 알려진 어떠한 바이러스와도 뚜렷한 상동성(60%)을 보이지 않아 WSBV가 기존의 바이러스와 구분되는 새로운 바이러스임을 알 수 있다. E3의 염기서열에 기초하여 한쌍의 PCR 프라이머를 작성하였다. 병든 새우로부터 분리한 DNA를 30회 증폭한 결과, 예상크기의 산물을 얻을 수 있어 이 방법은 바이러스의 감염여부를 알아낼 수 있는 진단법으로도 활용가능하다. Baculovirus associated with white spot syndrome (WSBV) is the causative agent of a disease with high mortalities and causes severe damage to shrimp cultures. In this study, we analyzed a recombinant clone (E3) obtained from a viral genomic library to characterize the causative agent in diseased shrimp Penaeus chinensis with white spot syndrome. According to the analysis of nucleotide sequence of E3, this clone did not showed considerable sequence homology with those of other known viruses, including baculovirus Autographa californica nuclear polyhedrosis virus (AcNPV), indicating that WSBV is a novel virus causing a serious disease in P. chinensis. Based on the sequence of E3 clone, a pair of PCR primers was designed. After 30 cycles of amplification, a specific product of the expected size was detected only if the total nucleic acids extracted from the diseased shrimp was used as a template DNA, suggesting that this method can be used to diagnose the virus infection in diseased shrimp.

Acinetobacter sp. BE-254에 의한 유화제의 생산

최경숙,장경립,임이종,김순한,정영기,이태호 동의대학교 기초과학연구소 1997 基礎科學硏究論文集 Vol.7 No.1

The strain producing bioemulsifier was isolated from soil samples. The isolated strain was identified as the genus Acinetobacter through its morphological, cultural and physiological characteristics. The highest emulsification activity and stability by Acinetobacter sp. BE-254 was observed after 5 days of cultivation in the culture medium containing n-hexadecane 4%, NaNO₃0.2%, KH₂PO₄0.01%, MgSO₄·7H₂O 0.01%, CaCl₂0.01%, and yeast extract 0.01%. The optimum pH and temperature for bioemulsifier production were pH 7.0 and 30℃, respectively. Furthermore the most of bioemulsifier was produced during the exponential growth phase, and this suggested that the bioemulsifier production was growth-associated. The bioemulsifier showed good emulsification activity on various emulsifying substrates such as hydrocarbons, edible oils, and petroleum fractions.

Cooperative stimulation of cisplatin-mediated apoptosis by hepatitis B virus X Protein and hepatitis C virus core Protein

권현진,장경립,Kwun, Hyun-Jin,Jang, Kyung-Lib Korean Society of Life Science 2007 생명과학회지 Vol.17 No.6

The co-infection with hepatitis B virus (HBV) and hepatitis C Virus (HCV) is associated with a more severe liver disease and increased frequency in the development of hepatocellular carcinoma com-pared to those with single infection. Here, we demonstrated that HBV X protein (HBx) and HCV Core cooperatively up-regulated the level of p53 in human hepatoma HepG2 cells. The elevated p53 subsequently stimulated the expression of proapoptotic Bax whereas it repressed the expression of antiapoptotic Bcl2. These effects, however, were not observed in p53-negative Hep3B cells. Consistently to their cooperative regulation of apoptotic effectors, HBx and HCV Core additively stimulated cisplatin-mediated apoptotic cell death of HepG2 but not of Hep3B cells. These results may help to explain the development of a more severe liver disease in patients co-infection with HBV and HCV as well as some contradictory results on the roles of HBx and Core in apoptosis. B형 간염 바이러스(HBV)와 C형 간염 바이러스(HCV)에 함께 감염되면 단독 감염의 경우보다 더 심각한 간질환이 유발되고 간암으로의 발전 가능성도 높아진다. 본 연구에서는 HBV의 X단백질(HBx)과 HCV의 코어 단백질이 인간 간암세포주인 HepG2세포에서 p53의 양을 협조적으로 증가시킨다는 것을 보여 주었다. 이로 인하여 세포예정사를 촉진하는 Bax 단백질의 발현이 더 증가하는 반면에 세포예정사를 억제하는 Bcl2의 발현은 더 억제됨이 관찰되었다. 그러나 이러한 효과들은 p53-음성인 Hep3B 세포에서는 관찰되지 않았다. 나아가 HBx와 코어 단백질은 HepG2의 cisplatin-매개성 세포예정사를 협조적으로 증가시키는 반면에 Hep3B에서는 이러한 효과가 나타나지 않았다. 이러한 연구 결과들은 HBV와 HCV가 동시에 감염되었을 경우에 나타나는 임상적인 소견을 이해하고 세포예정사에 미치는 HBx와 코어 단백질의 영향에 대한 기존의 상충적인 연구결과들을 해석하는데 도움을 줄 수 있다.

보리새우류의 바이러스 감염증의 진단

허문수,정초록,장경립 한국생명과학회 1999 생명과학회지 Vol.9 No.4

Baculovirus(WSBV) was isolated from infected Penaeide was collected from shrimp farm at southern sea of Korea from 1993 to 1995. The Infectious virus was purified and used for diagnosis of infected shrimp. Anti-viral serum were used for immunological detection as enzyme linked immunoabsorbent assay(ELISA) and indirect fluorescent antibody technique(IFAT). In IFAT, stomach, lymphoid organ and antenae gland of infected shrimp showed fluorescent reaction. In ELISA, tissues of spontaneously infected shrimp appeared higher O.D. values than in artificial infected shrimp. Primer set was constructed from sequence of 420bp of cloned Baculovirus(WSBV) genome. Specific band for infected shrimp was detected in Polymerase chain reaction(PCR)

2-O-α-D-Glucopyranosyl L-Ascorbic acid 생산을 위한 Cyclodextrin glucanotransferase의 고정화

성경혜,김성구,장경립,전홍기 부산대학교 김치연구소 2003 김치의 과학과 기술 Vol.9 No.-

AA를 AA-2G로 전환하는 CGTase를 고정화하여 AA-2G 대량 생산의 가능성을 고찰한 결과, CNBr-sepharose 4B가 가장 효과적인 담체로 판명되었고, CGTase의 고정화 최적 조건과 고정화 CGTase에 의한 AA-2G 생산의 최적조건 및 재사용성을 검토하였다. 최적 고정화 조건은 효소량 24,000 units/g resin으로 9시간 반응하여 약 18,000 units/g resin의 최고 고정화율을 얻을 수 있으며, pH 5.0(50 mM sodium citrate buffer)용액에 12% AA-Na와 8% soluble starch를 기질로 하여 800 units/ml의 고정화 CGTase를 첨가한 후 37℃에서 100 rpm으로 교반하면서 25시간 반응하여 약 18 mM의 AA-2G 최고량을 얻을 수 있었다. 또한 0.015 mM의 CaCl₂를 첨가하여 고정화 CGTase의 재사용성을 관찰한 결과, 5회까지 50% 이상의 AA-2G 생산율로서 그 가능성을 입증할 수 있었다. 그리고 효소의 재사용성이란 측면에서, 본 고정화의 다른 한 방법인 ultrafiltration(한외 여과)에 의해서는 Millipore사의 YM10 membrane을 이용하여 먼저 단백질량과 효소 활성 변화를 측정하여 그 가능성을 확보할 수 있었으며, 기질 20 ml을 사용하여 AA-2G를 생성시킨 후, 한외여과에 의해 효소만을 회수하여 연속 반응해 본 결과 8회까지 50% 의 생성률을 유지하였다. 따라서 한외 여과는 CNBr-sepharose 4B와 함께 효율적인 고정화의 한 방법으로 판명되었으며, 앞으로 이들 고정화 효소를 이용한 연속 반응 시스템의 구축이 뒤따라야 할 것이다. Cyclodextrin glucanotransferase (CGTase) from Paenibacillus sp. JB-13 was immobilized on various carriers by several immobilization methods such as ionic binding, covalent linkage and ultrafiltration to improve the process performance. The ultrafiltration and covalent linkage with CNBr-activated sepharose 4B were found as the best method for immobilization of CGTase. The ability of was as follows: contact time 6 hrs at 37℃, pH6.0, 100rpm and enzyme loading 24,000 units/g resin. The optimum conditions for production of 2-O-α-D-Glucopyranosyl L-Ascorbic acid by immobilized CGTase turned out to be: pH 5.0, temperature 37℃. 20% substrate solution containing 8%(w/v) of soluble starch and 12%(w/v) of L-ascorbic acid sodium salt, 100 rpm, for 25 hrs and with 800 units of immobilized CGTase/ml substrate solution. More over the CGTase activity could be stably maintained for 8 times of repetitive reactions after removing products by ultrafiltration through YM 10 membrane.

Phospholipase D activity is elevated in hepatitis C virus core protein-transformed NIH3T3 mouse fibroblast cells

김준모,최복희,장경립,민도식 생화학분자생물학회 2004 Experimental and molecular medicine Vol.36 No.5

Hepatitis C Virus (HCV) is associated with a severe liver disease and increased frequency in Overexpresion of HCV core protein is known to transform fibroblast cells. Phospholipase D (PLD) activity is comonly elevated in response to mitogenic signals, and has also ben overex-pressed and hyperactivated in some human cancer cels. The aim of this study was to understand how PLD was regulated in the HCV core protein-transformed NIH3T3 mouse fibro-blast cels. We observed that PLD activity was elevated in the NIH3T3 cels overexpresing HCV core protein over the vector alone-trans-of PLD protein and protein kinase C (PKC) in the HCV core protein-transformed cels was si-milar to the control cells. Phorbol 12-myristate 13-acetate (PMA), which is known to activate PKC, stimulated PLD activity significantly more in the core protein-transformed cels, in com-parison with that of the control cels. PLD activity assay using PKC isozyme-specific inhi-bitor and PKC translocation experiment showed that PKC-δ was mainly involved in the PMA- cells. Moreover, in cells overexpresing HCV core protein, PMA also stimulated p38 kinase more potently than that of the control cels, and an inhibitor of p38 kinase abolished PMA-in-duced PLD activation in cells overexpressing HCV core protein. Taken together, these results sugest that PLD might be implicated in core protein-induced transformation.