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Baculovirus를 이용한 Aujeszky's Disease Virus gⅢ 단백질 발현
송재영,이중복,현방훈,박종현,김병한,권창희,전무형,안수환 충남대학교 생물공학연구소 1993 생물공학연구지 Vol.3 No.-
The g Ⅲ gene located in U_L region of Yangsan strain, a field isolate of Aujeszky's disease virus (ADV) in Korea, was cloned into pTZ18R and sequenced. The gⅢ gene consisting of 1,437 nucleotides showed 98% sequence homology with that of Becker strain, a reference strain of ADV. The gene encoding gⅢ of Yangsan strain was placed under the control of Autographa californica nucleopolyhedrosis virus (AcNPV) polyhedrin promoter, and expressed by the derived recombinant baculovirus using Spodoptera frugiperda 9 (Sf9) cells. The expressed gⅢ was a protein with molecular weight of 72kd determined by immunoprecipitation and Western blotting assay using anti-ADV polyclonal antibodies and anti-gⅢ monoclonal antibody. The partially purified gⅢ protein was utilized as antigen in the radial immunodiffusion enzyme assay (RIDEA) to detect to specific antibody against ADV in pig sera. The results indicated that the sensitivity of RIDEA with the recombinant gⅢ protein antigen (98%) was as high as that with the conventional glycoprotein antigen extracted from the ADV infected cells. In addition, the false positive and false nagative reactions in gⅢ RIDEA were significantly reduced than the conventional glycoprotein RIDEA as judged from the results of standard serum neutralization test.
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.
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.
MLVA Genotyping of Brucella abortus isolated from cattle in Korea
Jin-Ju Lee(Jin-Ju Lee),Eun-Ji Yum(Eun-Ji Yum),Eun-Ji Park(Eun-Ji Park),Min-Kyeong Kim(Min-Kyeong Kim),Min-Hoe Lee(Min-Hoe Lee),So-Ra Sung(So-Ra Sung),Hee-Soo Lee(Hee-Soo Lee),Bang-Hun Hyun(Bang-Hun Hy 한국예방수의학회 2019 한국예방수의학회 학술대회자료집 Vol.2018 No.-
Culture supernatant proteins of Brucella canis represented potential antigenicity
Jin Ju Lee(Jin Ju Lee),So-Ra Sung(So-Ra Sung),Eun Ji Yum(Eun Ji Yum),Sung Dae Yang(Sung Dae Yang),Min Hoe Lee(Min Hoe Lee),Bang-Hun Hyen(Bang-Hun Hyen),Moon Her(Moon Her),Hee Soo Lee(Hee Soo Lee) 한국예방수의학회 2018 한국예방수의학회 학술대회자료집 Vol.2018 No.-
Phylogenetic classification of Brucella species based on whole genome sequencing
Eun Ji Yum(Eun Ji Yum),Jin Ju Lee(Jin Ju Lee),Se-Chul Kim(Se-Chul Kim),Sung Dae Yang(Sung Dae Yang),Min Hoe Lee(Min Hoe Lee),Bang-Hun Hyun(Bang-Hun Hyun),Moon Her(Moon Her),Hee Soo Lee(Hee Soo Lee) 한국예방수의학회 2018 한국예방수의학회 학술대회자료집 Vol.2018 No.-
Bang, Sunghee,Song, Ji Hoon,Lee, Dahae,Lee, Changyeol,Kim, Soonok,Kang, Ki Sung,Lee, Jong Hun,Shim, Sang Hee American Chemical Society 2019 Journal of agricultural and food chemistry Vol.67 No.7
<P>Roots of <I>Glehnia littoralis</I> have been used to heal stroke as a traditional medicine. Even though many studies on this plant have been conducted, the secondary metabolites produced by its endophytes and their bioactivities have not been investigated thus far. Therefore, a new meroditerpenoid named sartorypyrone E (<B>1</B>) and eight known compounds (<B>2</B>-<B>9</B>) were isolated from extracts of cultured <I>Neosartorya fischeri</I> JS0553, an endophyte of <I>G. littoralis</I>. The isolated metabolites were identified using spectroscopic methods and chemical reaction, based on a comparison to literature data. Relative and absolute stereochemistries of compound <B>1</B> were also elucidated. To identify the protective effects of isolated compounds (<B>1</B>-<B>9</B>) in HT22 cells against glutamate-induced cytotoxicity, we assessed inhibition of cell death, intracellular reactive oxygen species (ROS) accumulation, and calcium ion (Ca<SUP>2+</SUP>) influx. Among the isolates, compound <B>8</B>, identified as fischerin, showed significant neuroprotective activity on glutamate-mediated HT22 cell death through inhibition of ROS, Ca<SUP>2+</SUP> influx, and phosphorylation of mitogen-activated protein kinase, including c-Jun N-terminal kinase, extracellular signal-regulated kinase, and p38. The results suggested that the metabolites produced by the endophyte <I>N. fischeri</I> JS0553 might be related to the neuroprotective activity of its host plant, <I>G. littoralis</I>.</P> [FIG OMISSION]</BR>