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Construction of a live attenuated Salmonella strain expressing FanC protein to prevent bovine enterotoxigenic Escherichia coli and evaluation of its immunogenicity in mice
Gayeon Won , Hee Jung Kim , John Hwa Lee 대한수의학회 Korean Journal of Veterinary Research(구 대한수의학회지) 7 Pages
대한수의학회 Korean Journal of Veterinary Research(구 대한수의학회지) 2017, 제 57권 제 1호 2 9-15 (7 pages)
The expected size of the purified antigen was also confirmed via SDS-PAGE. The in-frame fusion of the rFanC was validated by nucleotide sequencing E. coli BL21 harbor- ing pET28a-K99 over-expressed rFanC (data not shown). Construction of a live attenuated Salmonella strain expressing rFanC The FanC protein (181 amino acids) was selected as a vac- cine candidate antigen against ETEC delivered by a... -
Production of transgenic pigs using a pGFAP-CreERT2/EGFP LoxP inducible system for central nervous system disease models
Seon-Ung Hwang, Kiyoung Eun, Junchul David Yoon, Hyunggee Kim, Sang-Hwan Hyun 대한수의학회 Journal of Veterinary Science 12 Pages
대한수의학회 Journal of Veterinary Science 2018, 제 19권 제 3호 13 434-445 (12 pages)
in the following sections. All other chemicals were purchased from Sigma-Aldrich (USA). Vector construction To construct pCDH-pGFAP-MCS-EF1-puro, the CMV promoter sequence of the pCDH-CMV-MCS-EF1-puro lentiviral vector (System Biosciences, USA) was removed by using two restriction enzymes, SnaBI and NheI. The pGFAP promoter sequence (17 upstream from the pGFAP transcription start site +1)... Transgenic (TG) pigs are important in biomedical research and are used in disease modeling, pharmaceutical toxicity testing, and regenerative medicine. In this study, we constructed two vector systems by using the promoter of the pig glial fibrillary acidic protein (pGFAP) gene, which is an astrocyte cell marker. We established donor TG fibroblasts with pGFAP-CreERT2/LCMV-EGFPLoxP and evaluated the effect of the transgenes on TG-somatic cell nuclear transfer (SCNT) embryo development. Cleavage... -
Establishment and evaluation of a murine ανβ3-integrin- expressing cell line with increased susceptibility to Foot- and-mouth disease virus
Wei Zhang, Kaiqi Lian, Fan Yang, Yang Yang, Zhijian Zhu, Zixiang Zhu, Weijun Cao, Ruoqing Mao, Ye Jin, Jijun He, Jianhong Guo, X 대한수의학회 Journal of Veterinary Science 8 Pages
대한수의학회 Journal of Veterinary Science 2015, 제 16권 제 3호 3 265-272 (8 pages)
bootstrap analysis using 1,000 replicates. Construction of the recombinant lentiviral plasmid The recombinant plasmid was constructed as previously described [29]. Briefly, pOK 12 and the internal ribosome entry site (IRES) fragment were amplified from pOK12 and the pIRES2-EGFP plasmid (stored in our laboratory), respectively, using the corresponding primer pairs (Table 1). The IRES fragment... Integrin ανβ3 plays a major role in various signaling pathways, cell apoptosis, and tumor angiogenesis. To examine the functions and roles of ανβ3 integrin, a stable CHO-677 cell line expressing the murine ανβ3 heterodimer (designated as “CHO-677-mανβ3” cells) was established using a highly efficient lentiviral-mediated gene transfer technique. Integrin subunits αν and β3 were detected at the gene and protein levels by polymerase chain reaction (PCR) and indirect... -
Molecular characterization of two Bangladeshi infectious bursal disease virus isolates using the hypervariable sequence of VP2 as a genetic marker
Md. Taohidul Islam*, Thanh Hoa Le, Md. Mostafizur Rahman, Md. Alimul Islam 대한수의학회 Journal of Veterinary Science 8 Pages
대한수의학회 Journal of Veterinary Science 2012, 제 13권 제 4호 10 405-412 (8 pages)
Biosystems, USA). Data analysis and phylogenetic construction For each IBDV strain, identical sequencingdata from two positive clones containing the expected VP2 DNA insert were recorded and analyzed. The sequences were edited with SeqEd v1.03 (Applied Biosystems, USA), aligned using AssemblyLIGN v1.9c (Accelrys, USA), and analyzed using the MacVector 8.2 package (Accelrys, USA) with a... Two Bangladeshi infectious bursal disease virus (IBDV) isolates collected in 2007, termed GB1 and GB3, were subjected to comparative sequencing and phylogenetic analyses. Sequence analysis of a 474-bp hypervariable region in the VP2 gene revealed that among four major amino acid substitutions observed in the strains, two were unique to GB1 and GB3 (Ser217Leu and Ala270Thr) while one substitution was only found in GB1 (Asn299Ser). Among IBDVs from Bangladesh including GB1 and GB3, the rate of... -
Establishment and characterization of an infectious cDNA clone of a classical swine fever virus LOM strain
Gil-Soon Park, Seong-In Lim, Seung-Ho Hong, Jae-Young Song* 대한수의학회 Journal of Veterinary Science 11 Pages
대한수의학회 Journal of Veterinary Science 2012, 제 13권 제 1호 12 81-91 (11 pages)
5 min extension at 68 oC. Construction of Flc-LOM To generate a full-length infectious cDNA clone of LOM, we used a modified methods previously described by Meyer et al. [16]. To assemble a single clone, the eight overlapping cDNA amplicons were inserted into a pACYC177 plasmid (New England Biolabs, USA), in which the kanamycin resistance site completely deleted, using available restriction... Classical swine fever virus (CSFV) causes a highly contagious disease among swine that has an important economic impact worldwide. CSFV strain LOM is an attenuated virus of low virulent strain of Miyagi isolated from Japan in 1956. Eight DNA fragments representing the genome of the CSFV strain LOM were obtained by RT-PCR. These were used to determine the complete nucleotide sequence and construct a full-length cDNA clone which was called Flc-LOM. Sequence analysis of the recombinant clone... -
Sequence Analysis of Canine LINE-1 Elements and p53 Gene in Canine Transmissible Venereal Tumor
Young-Ki Choi and Chul-Joong Kim 대한수의학회 Journal of Veterinary Science 8 Pages
대한수의학회 Journal of Veterinary Science 2002, 제 3권 제 4호 6 285-292 (8 pages)
a target cell for CAT assay and production of the virus like particles of SFV. Construction of plamids 5′truncated LINE-1 element of TVT (TVT-LINE) was subcloned into pBLCAT2 vector {PBL cat (F); TVT-LINE inserted into Xba I and BamH I sites upstream of CAT gene,PBL cat(P);TVT-LINE inserted into Sma I and Kpn I sites ofdownstream ofCAT gene,PBL cat(5D);5′deleted TVT-LINE which was digested with... LINEs (long interspersed nuclear elements or long interspersed repeated DNA elements) contains two open reading frames (ORFs), ORF1 and ORF2. We analysed the ORF2 located in the 5' region to the first exon of oncogene c-myc in canine transmissible venereal tumor (TVT) cell. We also showed the transcription activation was induced by this TVT-LINE sequence using CAT assay. To identify the mutation of tumor suppressor gene, sequence analysis of p53 from TVT cell was performed. We identified the... -
Differential Gene Expression in GPR40-Overexpressing Pancreatic β-cells Treated with Linoleic Acid
In-SuKim, So-YoungYang, Joo-HuiHan, Sang-HyukJung, Hyun-SooPark, Chang-SeonMyung 대한생리학회-대한약리학회 The Korean Journal of Physiology & Pharmacology 9 Pages
대한생리학회-대한약리학회 The Korean Journal of Physiology & Pharmacology 2015, Vol.19 No.2 9 141-149 (9 pages)
o C with 5% CO 2. Vector constructs For construction of the pCMV6-Neo-GPR40 expression plasmid, the vector pEZ-M02-GPR40 was purchased from GeneCopoeia (MD, USA) and in serted into the pCMV6-Neo vector (ORIGENE, MD, USA). E. coli DH-5α cells harbor- ing the selected plasmid DNA were cultured in LB broth media. Plasmid DNA was extracted using a plasmid mini kit (Qiagen, Hilden, Germany). The... “G protein-coupled receptor 40” (GPR40), a receptor for long-chain fatty acids, mediates the stimulation of glucose-induced insulin secretion. We examined the profiles of differential gene expression in GPR40-activated cells treated with linoleic acid, and finally predicted the integral pathways of the cellular mechanism of GPR40-mediated insulinotropic effects. After constructing a GPR40-overexpressing stable cell line (RIN-40) from the rat pancreatic β-cell line RIN-5f, we determined the... -
Roles of ERK and NF-κB in Interleukin-8 Expression in Response to Heat Shock Protein 22 in Vascular Smooth Muscle Cells
Seung-HunKang, Ji-HyukLee, Kyung-HaChoi, Byung-YongRhim, KoanhoiKim 대한생리학회-대한약리학회 The Korean Journal of Physiology & Pharmacology 6 Pages
대한생리학회-대한약리학회 The Korean Journal of Physiology & Pharmacology 2008, Vol.12 No.4 7 171-176 (6 pages)
luminometer and normalized to β- galactosidase activity. Construction of recombinant adenoviruses Recombinant adenoviral DNAs were prepared via homol- ogous recombination in E. coli, in accordance with AdEasyTM Adenoviral vector sy stem (Stratagene, La Jolla, CA). The transactivation domain deletion mutant of c-Jun (TAM67) (Brown et al. 1994) and the hemagglutinin-tagged inhibitor of NF-κ B... Heat shock proteins (HSPs) serve as molecular chaperones and play a role in cell protection from damage in response to stress stimuli. The aim of this article is to investigate whether HSP22 affects IL-8 expression in vascular smooth muscle cells (VSMCs), and which cellular factors are involved in the HSP-mediated IL-8 induction in that cell type in terms of mitogen activated protein kinase (MAPK) and transcription element. Exposure of aortic smooth muscle cells (AoSMCs) to HSP22 not only...