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Sequence Analysis of Canine LINE-1Elements and p53 Gene in Canine Transmissible Venereal Tumor
Chul-JoongKim,Young-Ki Choi 대한수의학회 2002 Journal of Veterinary Science Vol.3 No.4
LINEs (long interspersed nuclear elements or longinterspersed repeated DNA elements)contains two openreading frames (ORFs),ORF1 and ORF2.We analysedthe ORF2 located in the 5'region to the first exon ofoncogene c-myc in canine transmissible venereal tumor(TVT) cell.We also showed the transcription activationwas induced by this TVT-LINE sequence using CATassay. To identify the mutation of tumor suppressorgene, sequence analysis of p53 from TVT cell wasperformed. We identified the point mutation of 964nucleotide (T→C) resulting in the change of aminoacid (Phe→Ser) of p53 tumor suppressor protein.
Jie-YunPark,Sue-NiePark,Chul-JoongKim,HaryoungPoo,Hyun-MiPyo,Sun-WooYoon,Sun-YoungBaek 한국미생물학회 2002 The journal of microbiology Vol.40 No.4
To perform the prophylactic study of a vaccine derived from human papillomavirus (HPV) using Balb/c mice, we produced virus like particles consisting of HPV capsid protein L1 which has been reported to induce significant humoral and cellular immunity using various animal model systems. In order to produce HPV16 VLPs, the cDNA of L1 capsid protein in HPV type 16, obtained by polymerase chain reaction, was inserted into yeast expression vector, YEGa-HIR525 under the control of GAL10 promoter. The transformation of YEGa-HPV16 L1 was performed into the yeast Saccharomyces cerevisiae Y2805 by the lithium acetate method and the yeast clone expressing the highest level of L1 capsid protein of human papillomavirus type 16 was selected by Western blot analysis using anti-HPV16 L1 antibody. The purification of HPV16 VLP has been performed by the ultracentrifugation and gel-filtration methods. To validate the vaccine efficacy of the purified HPV16 VLPs and investigate the properties of HPV16 VLPs to induce humoral immunity, ELISA assay was performed. A significantly increased production of anti-HPV16 VLP antibodies was observed in sera from immunized mice. The neutralization activity of antibodies in the sera from the vaccinated mice was demonstrated by a rapid and simple assay to detect hemagglutihation inhibition activity.
Hyun-Soo Kim,In-WookHwang,Su-MiKim,Chul-JoongKim,Kwang-SoonShin,Hyun-SooKim 대한수의학회 2002 Journal of Veterinary Science Vol.3 No.1
The ORF5 gene ncodes a major envelopeglycoprotein (GP5), which is one of the thre majorproteins of porcine reproductive and respiratorysyndrome virus (PRRSV). The GP5 protein has benknown to be a 24.5-26kDa N-glycosylated envelopeprotein. The GP5 is involved in inducing neutralizingantibodies. For this reason, the GP5 is primary can-didate for the PRRSV subunit vaccine. To produce thecloned t h e O R F 5 g e n e f r o m P R S V C N V - 1 i n t o t h eSemliki Forest virus (SFV)-based expresion vector,resulting in recombinant pSFV-ORF5. By the infectionwith recombinant pSFV-ORF5 to BHK-21 cells, theGP5 expresion was confirmed by imunocytochemistryand imunobloting asay. The recombinant virusparticle harboring ORF5 gene was infectious to BHK-21and MARC-145. The RNA synthesis and expression ofGP5 in the infected cell was also confirmed byRT-PCR.
Hyun-Soo Kim,Tae-UkHan,Shien-Young,Kwang-SoonShin,Chul-JoongKim,Jong-TaikKim,Hyun-SooKim 대한수의학회 2002 Journal of Veterinary Science Vol.3 No.3
During the period from January to December of2001, a total of 3,391 swine sera were submited to urlaboratory from 256 farms for the diagnosis of porcinereproductive and respiratory syndrome (PRS). Theantibody to porcine reproductive and respiratorysyndrome virus (PRRSV) was tested by the indirectimunofluorescent antibody (IFA) test. Of the 256farms tested, 230 farms (89.8%) were positive for thePRRSV antibody was 52.1% (1765/3391). Most of thepigs eemed to be infected with PRSV at around 50to 60 days old. The seroprevalence of the antibodybecame higher with age, and peaked at around 100days old. More than one-third of the adult pigs,including boars, gilts, and sows, was positive for thePRSV antibody. The infection of PRSV was chronicand confined to growers and/or finishers in mostfarms. However, the antibody was detected in alproduction phases at some farms .