• Journal of Life Science
• /
• v.15 no.2 s.69
• /
• pp.192-196
• /
• 2005

The SNF2/SW12 family comprises proteins from a variety of species with in vivo functions, such as transcriptional regulation, maintenance of chromosome stability during mitosis, and various types of DNA repair. This study was shown the characterization of hrp2+ gene which was isolated by PCR amplification using the conserved domain of SNF2 motifs. Sequence analysis of hrp2+ gene showed striking evolutionary conservation among the SNF2 family of proteins. The transcript of hrp2+ gene was found to be a 4.7 kb as identified by Northern hybridization. To investigate the inducibility of hrp2+ gene, transcript levels were examined after treating the cells to various DNA damaging agents. The transcripts of hrp2+ were induced by UV-irradiation. But the transcripts were not induced by treatment of $ 0.25\%$ Methylmethane sulfonate (MMS). These results implied that the effects of damaging agents are complex and different regulatory pathways exist for the induction of this gene. Hrp2 protein was purified near homogeneity by combination of affinity chromatography. We tested the purified Hrp2 protein for the helicase activity in an oligonucleotide release assay. However we were unable to detect any helicase activity associated with the Hrp2 protein, indicating that the helicase motifs in Hrp2 are merely indicators of a broader DNA-dependent ATPase activity.

• Journal of Veterinary Science
• /
• v.21 no.5
• /
• pp.64.1-64.14
• /
• 2020

Background: Canine distemper virus (CDV) infection results in high morbidity and mortality in dogs. There has been no report about Isolation of Korean CDV since 1980 in Korea. Objectives: To investigate the biological properties and the genetic characterization of Korean CDV. Methods: Vero cells expressing dog signaling lymphocyte activation molecule (dSLAM) gene named as Vero/dSLAM were used to isolate CDV using 17 samples. Diagnostic methods such as cytopathic effects, immunofluorescence assay, peroxidase linked assay, electron microscopy, rapid immunodiagnostic assay, and reverse transcription polymerase chain reaction were used to confirm the Korean CDV isolate as a CDV. The genetic analysis was performed through cloning and sequencing of hemagglutinin gene of CDV isolate. Results: A virus propagated in Vero/dSLAM cell was confirmed as CDV (CD1901 strain) based on the above methods. The CD1901 strain showed the highest viral titer (10^5.5 50% tissue culture infectious dose [TCID₅₀]/mL) in the Vero/dSLAM cells at 4 days post inoculation, but did not form a fork on chorioallantoic membrane of 7-day-old egg. Ribavirin, a nucleotide analogue anti-viral agent, inhibits moderately the Korean CDV propagation in the Vero/dSLAM cells. The nucleotide and amino acid sequences of the H gene of CD1901 strain were compared with those of other CDV strains. The CD1901 strain belonged to Asia 1 group and had the highest similarity (99.9%) with the BA134 strain, which was isolated in China in 2008. Conclusions: We constructed successfully Vero/dSLAM and isolated one Korean CDV isolate (CD1901 strain) from a naturally infected dog. The CD1901 strain belonged to Asia 1 genotype.

• Microbiology and Biotechnology Letters
• /
• v.17 no.6
• /
• pp.600-605
• /
• 1989

For the purpose of isolation of the Zymomonas mobilis DNA fragments showing promoter activity in Escherichia coli, a promoter screening vector, PCMT215 was constructed by transferring a promoterless chloramphenicol acetyltransferase (CAT) gene of pYEJ001 into pMT21 which contains $\beta$-lactamase gene and multiple cloning sites. A library of Z, mobilis Sau3AI DNA fragments was constructed in E. coli using the newly constructed pCMT215. Fourteen clones showing resistance to chloramphenicol ranging in concentration from 30 to 750 $\mu$g/$m\ell$ were selected. From five clones of them, the Z. mobilis DNA fragments expressing CAT gene of the recombinant plasmids were sequenced and then sites of transcriptional initiation were identified. The nucleotide sequences of the cloned DNA shared AT rich regions, poly A's or T's stretches and palindromic regions. The positions of transcriptional initiation for CAT gene occurred at more than one site spaced over by 4 to 190 base pairs on the cloned fragments in E. coli.

• Korean Journal of Veterinary Service
• /
• v.34 no.1
• /
• pp.45-54
• /
• 2011

This study was conducted to investigate the distribution of Salmonella spp. from pigs and cattle in slaughterhouse, the antimicrobial resistance pattern and the prevalence of resistance genes of isolates. A total of 640 fecal samples from pigs and cattle in slaughterhouse were collected for isolation of Salmonella spp.. Isolation rate was revealed as 15% in pigs and 1.6% in cattle. As result of serotyping, group B (56.6%) were identified as most common in pigs and cattle isolates, in order of group C (24.5%) and group E (15.1%). S. Typhimurium (50.9%) was most common serotype. The major serotypes were in order of S. Rissen and S. London (11.3%) and S. Riggil (7.6%). In antimicrobial test, all isolates were demonstrates susceptibility to nitrofurantoin. But isolates were revealed resistance other antibiotics in order of tetracycline (64.6%), streptomycin (68.3%), ampicillin and amoxicillin (56.3%) and spectinomycin (47.9%). With polymerase chain reaction, antimicrobial resistance gene strA (75.0%) and aadA1 (3.1%) were detected in streptomycin resistance isolates and tetA (94.3%) and tetB (11.3%) gene were detected in tetracycline resistant isolates, but tetG was not detected. Class 1 integron gene was detected in all Salmonella isolates.

• Journal of Animal Science and Technology
• /
• v.46 no.2
• /
• pp.155-164
• /
• 2004

This involves identifying and cloning trapped genes from cultured cells carrying the gene-trap constructs and generating cloned zebrafish using these cells for functional study. Gene-trapping studies in gene-trapped cells were carried out in initial and cloned zebrafish carrying gene-trap events were successfully produced based on the nuclear transplantation technique. Two kind of retroviral gene-trap constructs were adopted. The first one(SA/GFP-TP), constructed in my laboratory, carries a GFP reporter gene containing a splicing acceptor and an internal neo gene. The second one(Neo-TP), obtained from Dr. Hicks (Hicks et al., 1997), contains a promoter-less neo gene located in the LTR sequence of a retroviral vector. The infected cells were subjected to drug selection(neomycin treatment) because the two constructs carry the neomycin resistant gene. All those cells survived the neomycin treatment should carry the proviral insertions. For Neo-TP, Isolated DNA from the neomycin-resistant fibroblast cells infected by Neo-TP, was digested with EcoR1 restriction enzyme and transformed into bacteria after ligation. This procedure led to the isolation of seven clones carrying flanking cellular DNA with a typical retroviral integration signature sequence. These clones contained genomic DNA ranging from 1kb to 7kb and sequences of 300-600 bp were obtained from each of the rescued plasmids. Database searching showed that all of them share high homology to zebrafish sequences. For fish cloning using tagged cells, initially, nucleus donors directly selected from a mixture of cells(Neo-TP cells) were used. A total of 44 embryos(3.7%) out of 1179 transplants were reached blastula stage; 8 of these embryos(0.8%) hatched and 3(0.3%) of them survived to adulthood. One out of three lived cloned zebrafish has an amplified fragment and was labeled with 32P.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 2003.10a
• /
• pp.83.2-83
• /
• 2003

A full length cDNA, OsLEUZIP, encoding leucine zipper containing protein from rice EST of rice (0ryza sativa L. cv. Dongjin) treated Xanthomonas oryzae pv. oryzae 10331. OsLEUZIP contains 1,227 bp nucleotides and encodes a protein of 408 amino acid residues with predicted molecular weight of 47,229 Da. The deduced amino acid sequence of OsLEUZIP has consensus sequence of leucine zipper from PROSITE (PDOC00029), L-X(6)-L-X(6)-L-X(6) -L. OsLEUZIP gene were preferentially induced in rice during incompatible interaction with Xanthomonas oryzae pv. oryzae 10331 and Pyracuraria grisea KJ-301. Expression of OsLEUZIP gene was also induced by treatment of abiotics such as ethephon and ABA. Our data represented in this study suggesting that OsLEUZIP gene may play an important role in the rice defense-related. Further studies of this gene, overexpression in rice, yeast-two hybrid assay, electrophoretic mobility shift assay and northern blot analyses of transgenic plant, would be useful to elucidate the role of the OsLEUZIP gene in defense responses of rice.

• Korean Journal of Plant Tissue Culture
• /
• v.22 no.3
• /
• pp.149-155
• /
• 1995

The coat protein (CP) gene was cloned from RNA genome of the Cucumber Mosaic Virus strain ABI (CMV-ABI) isolated in Korea. The comparisons of the nucleotide sequence of the cloned CP gene and its deduced amino acid sequences with other CP genes revealed that the CMV-ABI belongs to subgroup I (type I), CMV-ABI developed the typical mosaic symptom in infected plants. Tobacco plants (Samsun and NC82) were transformed by leaf-disc transformation via Agrobacterium, temefaciens LB4404 harboring pVCP, witch CMV-ABI CP gene was inserted into the pBI121, and a number of mature transgenic tobacco plants were developed. Southern and PCR analysis of genomic DNA from the transgenic plants showed that the CP gene was integrated into the genomes of the most of the transgenic plant. Result of the segregation patterns of resistance in T1 seedlings of the plants to kanamycin showed that the transgenic plants containing l,2 and 3 copies of CP gene were50%, 39% and 11% of the total transgenic plants, respectively.

• Korean Journal of Plant Tissue Culture
• /
• v.28 no.5
• /
• pp.255-261
• /
• 2001

Vitis vinifera thaumatin-like protein (VVTL1) is a fruit-specific and ripening-related protein in grape. In order to isolate VVTL1-homolog gene and fruit-specific promoter from Campbell cultivar, we isolated a genomic clone containing VVTL1-homolog gene from grape genomic library through plaque hybridization. VVTL1-homolog gene has an intronless genomic structure, which the pattern is matched with those of other PR5 genes such as osmotin and osmotin-like protein genes. Transcription start site was determined by primer extension analysis. The promoter region of VVTL1-homolog gene contains a sequence or structure, especially the location and number of TCA box and ABRE (abscisic acid-responsive element), distinct from other reported plant PR5 genes, though with several known functional elements such as a TATA box and CAAT box. These results suggested that VVTL1-homolog gene may be regulated by a plant hormone, abscisic acid, and one or several stresses such osmotic pressure and pathogen infection. The isolation of fruit-specific promoter may be helpful to breed a genetically modified grape with valuable phenotype or materials in fruits.

• Korean Journal of Microbiology
• /
• v.44 no.3
• /
• pp.251-257
• /
• 2008

Four potential microcystin-producing cyanobacteria were isolated from large reservoirs which act as sources of drinking water supply in Korea. Strain DC-2, YD-l, and YD-6 were closely related to Microcystis aeruginosa based on the analysis of l6S rRNA gene and mcyA gene sequences. mcyA gene sequence of YDS2-3, isolated from Yongdam Reservoir, was closed to that of M. aeruginosa, whereas l6S rRNA gene sequence was not related to the known sequences of microcystin-producing cyanobacteria indicating this strain can be a novel cyanobacterium belonging to the genus Microcystis. When mcyA gene sequences of isolated cyanobacteria were compared with the mcyA gene sequence library of two reservoirs, the sequence of DC-2 matched with the dominant ones.

• Korean Journal of Food Science and Technology
• /
• v.35 no.3
• /
• pp.470-474
• /
• 2003

DNA sequence information on small-subunit rRNA gene (16S rDNA) obtained from food-poisoning bacterial culture was used to investigate the presence of bacterial pathogens in food. By reverse dot blot detection method, presence of food-poisoning bacteria could be confirmed on hybridization of digoxigenin-labeled 16S rDNA Polymerase Chain Reaction (PCR) primer product and biotin-labeled specific oligonucleotide probe. Escherichia coli, Bacillus cereus. and Salmonella sp. were used as the representative food-poisoning bacterial microorganisms. An oligonucleotide probe, based on the variable region of 16S rRNA gene, was used as the specific probe. These tools may be more useful than classic biochemical method for rapid identification of contaminated food.