Baculovirus Vector System에 의해 발현된 재조합 Pseudorabies Virus Major Capsid Protein의 면역원성

전무형,안동준,장경수,조용성,박종현,송재영,현방훈,안수환 충남대학교 생물공학연구소 1997 생물공학연구지 Vol.5 No.-

The recombinant pseudorabies virus major capsid protein (rMCP) was produced by expression of the MCP gene in Sf-9 cell using baculovirus transfer vector system. Following evaluation of the immunochemical properties of the rMCP, the immunogenicity of the recombinant subunit protiens were investigated in guinea pig and swine to obtain the preliminary guide line for the subunit vaccine using rMCP and gP50. It was proved that ultrasonication and 30% ammonium sulfate was most efficient to concentrate and purify the protein. The rMCP was safe in mice, guinea pigs and piglets. In guinea pigs, rMCP mixed with various adjuvants induced substantial degree of serum neutralizing antibody titers, but revealed incomplete protecivity against challenge. In swine, the combination of rMCP and gP50 showed the higher serum neutralizing antibody titers and cellular immune responses than rMCP alone. However, the protectivity was lower in comparison with the commercial gI-deleted inactivated vaccine. We expect these results to contribute to characterization of MCP gene of Korean isolate of PRV and to ultilize as preliminary information for prodution and evaluation of PRV recombinant subunit vaccines.

Message Filtering을 통한 TMN의 성능향상에 관한 연구

이종옥,김현준 동국대학교 경영대학원 1996 經營論叢-東國大學校 經營大學院 Vol.20 No.-

Today, Distributed Telecommunication Networks are consist of various hetrogeneous equipments. ITU-T suggests TMN for managing these hetrogeneous equipment and services. In reality, there is a few equipment which support ITU-T suggestions. So Q Adaptor is in charge of converting non-TMN qiupment's interface into TMN's. But these QAF(Q Adapter Function) can cause significant overhead if there are many non-TMN equipments in the network. In this paper, the method which can improve management efficiency in TMN by reducing QAF's overhead is proposed. By moving Q Adapter's filtering function from TMN's management system to the outside, the QAF's overhead can be reduced. Prototype network Management System was built using NetExpert, lex and c. The proposed system was simulated using 1,647,977 sample messages from TDX-PS switches. The result of the simulation are as follows; 1) With proposed system, the number of message row was reduced from 9,876,917 to 6,067,913 with 61.44 % reducing rate. 2) With proposed system, Q Adapter's message processing time was reduced from 4,961 seconds to 1,471 seconds with 29.7% reducing rate. 3) With proposed Message Filtering System, the QAF's performance improvement was founded, so TMN's management efficiency is expected. Though there is significant improvement on Q Adaptor's functions, the proposed system has some limitaions. The prototype was developed and simulated in batch processing mode and for only TDX-PS switches. So, from now on, the sudy on generalizing proposed system to fit on only hetrogeneous equipment will be suggested.

Pocine Adenovirus-3의 E1B Region의 鹽基序列 分析

朴鍾賢,宋載永,李重馥,玄芳勳,安東濬,車相昊,裵用泰,姜永源,Reddy, P S,全茂炯,安壽煥 충남대학교 생물공학연구소 1999 생물공학연구지 Vol.7 No.-

돼지 아데노바이러스(PAV-3). 6618주의 EIB region이 包含되어 있는 map unit 4.0에서 9.7까지의 유전자에 대한 1,984 bp의 염기서열을 決定하였으며, 이 結果를 알려진 여러 아데노바이러스 유전자와 비교하여 다음과 같은 결과를 얻었다. 1. PAV-3의 EIB유전자는 10개의 ORF로 구성되어 있으며, 그 중 아데노바이러스의 단백질과 유사성이 있는 것은 ORF1, ORF2 및 ORF3이었다. ORF1은 Ad41의 19kd 와 BAV-2에서의 EIB ORF2에서의 아미노산의 一致率은 각각 32%와 31%이었다. 2. ORF2는 Ad2 55kd protein과 tupaia adenovirus 44kd protein가 각각 34%로 아미노산 一致率이 가장 높았으며, Ad41의 52kd protein. BAV-3의 EIB ORF3에서도 33%의 一致率을 보였다. 3. ORF1은 61-666 uncleotide (606 bp), ORF 2에서는 429-1,850 uncleotide (1,422 bp)의 부위로 각각 202, 474 a.a로 構成되었으며, 예상되는 분자량은 20 kd와 52 kd이었다. 4. ORF3는 hexon-associated pIX유전자로 추정되며 내부에 1개의 polyadenylation signal(ATAAA)이 1938-1942 uncleotide에 위치하였으며, 이 부위는 TATA box (1937-1942 uncleotide)와 중복되어 존재하였다. Porcine adenovirus type 3 (PAV-3) does not cause severe infection in pigs. Adenovirus has been suggestive of live vaccine vector carrying foreign gene. One of insertion regions is delayed early (EIB) region. However, EIB region of PAV-3 has not been molecularly characterized to date. Nucleotide sequence of EIB of PAV-3 was determined. The EIB region was composed of 1,984 bp and located between 4.0 and 9.7 map units. Three potential open reading frames(ORFs) with low level of homology to other adenoviruses and a polyadenylation signal were identified in the rightward direction of genome. The nucleotide and the predicted amino acid sequences of EIB were compared to those of human and animal adenoviruses. One of the three potential ORFs. ORF1 encoded a polypeptide homologous to bovine adenovirus type 2(BAV-2) ORF2 and human adenovirus type 41(Ad41) 19 kd protein. ORF2 encoded a polypeptide homologous to human adenovirus type 2(Ad2) 55 kd protein, bovine adenovirus type 3(BAV-3) ORF3 and porcine adenovirus type 4(PAV-4) ORF2. The predicted protein of ORF1 had homology to those of Ad41 and BAV-2 with 32 and 31% respectively, whereas the deduced protein of ORF2 had homology to those of Ad2. BAV-3 and PAV-4 with 34, 33 and 29%, respectively.

외안각 절제술과 경결막 절개에 의한 안와 접근법

정종철,김성범,서지훈,송민석,전창훈,최세훈,김현민 대한악안면성형재건외과학회 2002 Maxillofacial Plastic Reconstructive Surgery Vol.24 No.4

Various surgical approaches to the orbits have been used. But the transconjunctival approach, also called the inferior fornix incision, provides adequate access to the orbital floor with esthetic results. Additionally, transconjunctival approach with lateral canthotomy allows more wide surgical field and good access to the orbital area. Generally, preseptal approach is more used rather than postseptal approach in the transconjunctival incision for the prevention of fat herniation during surgery. There are some complications in transconjunctival approaches but these complications can be prevented with careful surgical techniques. We report clinical results in 12 patients who were treated by preseptal transconjunctival incision with lateral canthotomoy for the treatment of orbital trauma. We were satisfied with this technique for reduction and reconstruction of the orbital trauma and we could also observe relatively good esthetic results.

Pseudorabies Virus의 Major Capsid Protein 유전자의 클론닝과 Baculovirus Vector System에 의한 발현

안동준,전무형,송재영,박종현,현방훈,장경수,안수환 충남대학교 생물공학연구소 1997 생물공학연구지 Vol.5 No.-

Pseudorabies is caused by Pseudorabies virus (PRV: Aujeszky's disease virus) of Herpesviridae that is characterized by 100 to 150nm in size with a linear double-stranded DNA molecule with of approximately 90 × 10 exp (6)Da. This disease affects most of domestic animals such as swine, cattle, dog, sheep, cat, chicken, etc. causing high mortality and economic losses. In swine, young piglets show high mortality and pregnant sows, reproductive failures. However the adult swine reveals no clinical signs in general. But they become a carrier state and play an importnat role for propagation of the disease. In this study, the nucleotide sequence of major casid protein gene of PRV, Yangsan strain isolated from the diseased swine in Korea was analyzed, and the recombinant MCP was produced by expression of the MCP gene in Sf-9 cell using baculovirus transfer vector system. As result, in BamHI digestion, MCP gene locus of PRV YS strain showed different from that of Indiana S strain. The patterns of enzyme mapping were also found to be unidentical each other. The sequence of the MCP gene partially analyzed showed 98.09% identity to Indiana S strain. The expression of MCP in Sf-9 cell cotransfected by pVLMCP-44 baculovirus expression vector was characterized by Southern blot hybridization, immunofluoresent and immunocytochemical tests, SDS-PAGE and Western blotting. The rMCP with M.W. 142kDa was most effectively expressed in Sf-9 cells at the 3-4th days post inoculation of the recombinant baculovirus by 2 moi.

국내에서 분리된 canine parvovirus의 구조유전자 cloning과 염기서열 분석

박종현,송재영,이중복,현방훈,안수환,전무형 충남대학교 생물공학연구소 1993 생물공학연구지 Vol.3 No.-

In this study gene encoding structural proteins of a CPV isolate was cloned and saquenced to elucidate the molecular genetical properties of the canine parvoviruses isolated from the field. Six recombinant plasmids of pEP3, p1471, p2070, pEP069, pEP338 and p14711p were constructed from the map positions 22 to 98 of RF DNA to clone the VP1 and VP2 genes of CPV-V20. Sequentialy the gene comprising 3780 nucleotides were sequenced by dideoxy chain termination method. When nucleotide sequence of gene encoding the structural proteins of CPV-V20 was compared with those of other strains, CPV-N, CPV-d and CPV-780929 published previously. DNA homologies to CPV-V20 were 99.87% with CPV-NM, 99.73% with CPV-d, 96.85% with CPV-780929 AND 98.4% with FPLV-Carl, respectively. The DNA sequence data of CPV-V20 showed seven point mutations and also deletion of 135 nucleotides from the nucleotide position 4745 to 4879 located in the 3-noncoding region of CPV-N.

좌심실비후 환자에서 베타차단제의 급중단이 임상증상 및 좌심기능에 미치는 영향

황종현,김영태,배용학,임현주,조용근,채성철,전재은,박의현 慶北大學校 醫科大學 1997 慶北醫大誌 Vol.38 No.1

목적 : 베타 차단제와 칼슘 길항제의 장기투여로 안정상태에 있는 비후형 심근증이나 고혈압성좌심실비후 환자에서 이들 약제를 갑자기 중단하였을 때 혈역학, 심기능 및 임상상에 미치는 영향을 알아보기 위하여 본 연구를 시행하였다. 대상 및 방법 : 경북대학교 병원 순환기내과에서 통원치료중인 비후형 심근중 환자 5명과 심한좌심실비후를 동반한 고혈압 환자 6명(남자 6명, 여자 5명)을 대상으로 장기투여 중이던 베타 차단제와 칼슘 길항제를 최소 5일 중단한 후 임상증상, 혈압 및 심박수의 변동, 그리고 투약중지 전후에 도플러 심초음파도를 기록하여 좌심실내경, 벽두께, 좌심실근량, 구혈율 및 승모판혈류속도곡선의 변화를 관찰하였다. 결과 : 1) 약물치료 중단 후, 베타차단제의 경우 9명 중 6명에서, 그리고 칼슘길항제 및 디소피라미드를 복용하던 각 1명, 도합 7명의 환자에서 심계항진, 호흡곤란의 악화, 흉통이 관찰되었고, 심계항진을 호소한 1예에서는 심방세동이 재발되었다. 2) 혈압, 심박수, 확장말기 및 수축말기 좌심실내경, 심실중격 및 심실후벽 두께, 좌심실근량 및 구혈을 모두는 치료약물 중단 전후에 다소 변화하였으나 유의하지 않았다. 3) E파 최고속도와 E파 시간속도적분은 약물투여 중단 전후에 유의한 변화는 없었으나, A파 최고속도와 A파 시간속도적분은 중단 후에 각각 유의하게 증가하였으며(both P<0.01), E/A 최고속도비와 E/A 시간속도적분비는 각각 유의하게 감소하였다(both P<0.01). 한편, E파 감속시간, 등용이완시간 그리고 A-Ar 간격[A파의 최고점에서 A reversal(Ar)파의 최저점간의 시간] 모두는 중단 후 유의한 변화가 없었다. 1 결론 : 비후형 심근증 이나 고혈압성 좌심실비후 환자에서 장기투여중이던 베타 차단제와 칼슘길항제를 일시적으로 중단하였을 때 임상중상의 악화와 더불어 확장기 기능의 변화를 초래하였다. 따라서 약물치료로 비록 임상적으로 안정상태에 있더라도 치료를 갑자기 중단할 때는 주의해야 할 것으로 생각된다. Objectives : This study was done to evaluate the changes in clinical findings, hemodynamics and cardiac function after abrupt withdrawal of longterm beta-blockers and/or calcium channel blockers in hemodynamically stable patients with hypertrophic cardiomyopathy and left ventricular(LV) hypertrophy associated with hypertension. Methods : Eleven patients with hypertrophic cardiomyopathy(n=5) and LV hypertrophy associated with hypertension(n=6) were studied. Symptom assessment, blood pressure, electrocardiography and echocardiography including pulsed Doppler examination were obtained before and at least 5 days after abrupt cessation of beta-blockers and calcium antagonists. Results : 1) Deterioration in symptoms(i.e. palpitation, aggravation of dyspnea and chest pain) was observed in seven patients after abrupt withdrawal. 2) There were no significant changes in blood pressure, heart rate, end-systolic and end-diastolic LV dimensions, interventricular septal thickness, LV posterior wall thickness, LV mass, and ejection fraction before and after drug withdrawal. 3) Doppler time intervals and E wave peak-velocity and time velocity integral(TVI) were not significantly changed before and after drug withdrawal. But A wave peak-velocity and TVI after abrupt withdrawal were significantly increased(both p<0.01), and E/A peak-velocity ratio and TVI ratio were significantly decreased(both p<0.01). Conclusions : These findings suggest that temporary omission of beta blockers and calcium channel blockers develops deterioration in symptoms and LV diastolic dysfunction in hypertrophic cardiomyopathy and LV hypertrophy associated with hypertension. Physicians should avoid abrupt withdrawal of longterm beta blockers in LV hypertrophy.

Baculovirus를 利用한 Canine Parvovirus VP2蛋自質의 發現

朴鍾賢,宋載永,玄芳動,安動濬,姜永源,全茂炯,安壽煥 충남대학교 생물공학연구소 1998 생물공학연구지 Vol.6 No.-

국내에서 分離된 개파보바이러스주(V20주)의 VP2 遺傳子를 baculovirus system을 이용하여 발현시켜 다음의 結果를 얻었다. 1. 개파보바이러스의 VP2 유전자를 PCR에 의해 增幅하여 1755bp의 VP2遺傳子를 pUC19에 클로닝하여, 클로닝된 遺傳子를 polyhedrin promoter를 가지는 baculovirus expression vector인 pVL1393에 옮겨 VP2 발현벡터인 pVL1393-VP2를 얻을 수 있었다. 2. pVL1393-VP2 plasmid와 baculovirus DNA와의 homologous recombination에 의해 재조합바이러스인 VP2-BV를 얻을 수 있었으며, 그 발현효율은 2.000-5.000 HAU/0.05 ml이었다. 3. 免疫沈澱法에 의해 발현된 단백질은 개파보바이러스의 VP2단백질과 유사한 64 kb에 달하는 것이었으며, 血球凝集能을 지니고 있었다 4. 血球凝集能을 지닌 蛋白質이 여러 陽性血淸에 있어서 抗體수준을 測定할 수 있는지 개파보바이러스항원과의 相關性을 比較한 바 0.94 (n=125. p<0.01)의 相關係數를 보였다 5. 발현 VP2白은 virus-like particles를 形成하였으며, 그 크기로는 개파보바이러스와 비슷한 25 ㎚의 크기를 갖았다. Canine parvovirus(CPV) is a member of autonomous replicating parvoviruses and is aetiologically associated with enteritis and mycoarditis in puppies. The capsids of CPV are composed of three structural proteins: VP1, VP2 and VP3. The VP2 protein is the major component of capsid. The VP2 gene of a canine parvovirus. V20 strain isolated in Korea was cloned into baculovirus expression vector, and subsequently the VP2 protein was expressed by a recombinant baculovirus under the control of polyhedrin promoter. The recombinant VP2 protein expressed in Spodoptera frugiperda 9(Sf9) cells was detected by haemagglutination(HA) test and immunofluorescent antibody assay. Molecular weight of the recombinant VP2 protein expressed was estimated as 64Kd when tested by immunoprecipitation test using anti-CPV monoclonal antibody. In haemagglutination inhibition(Ⅲ) test. 8 HA units of the recombinant VP2 protein antigen was successfully utilized to determine a level of antibody against CPV in various positive sera. The recombinant VP2 protein showed also the capability to form virus like particles similar in size and appearance to the CPV virions.

소득작물 전후작지 적응 극단기성 벼 ‘중모 1032’

손영보,이종희,조준현,이지윤,오성환,송유천,박동수,박노봉,김상열,여운상,남민희,권영업 忠北大學校 農業科學硏究所 2015 農業科學硏究 Vol.31 No.2

꿩에서 분리된 Newcastle Diesase Virus 내열성주 (CBP)의 Fusion(F) 유전자 클론닝과 염기서열 분석

장경수,전무형,송희종,김귀현,박종현 충남대학교 생물공학연구소 1999 생물공학연구지 Vol.7 No.-

The gene encoding F protein of CBP-1 strain, a heat-stable Newcastle disease virus (NDV) isolated from the diseased pheasants in Korea, was characterized by reverse transcriptionpolymerase chain reaction (RT-PCR), nucleotide and amino acid sequences. Virus RNA was prepared from the chorioallatoic fluid infected with NDV CBP-1 virus and cDNA was amplified by RT-PCR, cloned and sequenced to analyze. The PCR was sensitive as to detect the virus titer above 2^5 hemagglutination unit. 1.7kb (1,707bp) size of the cDNA was amplified and cloned into BamHI site of pVL1393 Baculo transfer vector. The uncleotide sequences for F protein were determined by dye terminator cyclic sequencing using four pairs of primers, and 553 amino acid sequences were predicted. In comparison of the nucleotide sequence of F gene of CBP-1 with those of other NDV strains, the homology revealed 88.8%, 98.5% and 98.7% with Kyojungwon (KJW), Texas GB and Beaudette C strains, respectively. As the deduced 553 amino acid sequences of F protein of CBP-1 were compared with those of other NDV strains, the homology appeared 89.9%, 98.7% and 98.9% with KJW, Texas GB and Beaudette C strains, respectively. The putative protease cleavage site (112-116) was R-R-Q-K-R, indicating that CBP-1 strain is velogenic type. The amino acid sequences include 6 sites of N-asparagine-linked glycosylation and 13 cysteine residues. These data indicate that the genotype of CBP-1 strain is more closely associated with the strains of Texas GB and Beaudette C than KJW strain.