• 한국한의학연구원논문집
• /
• 제1권1호
• /
• pp.477-493
• /
• 1995

In order to facilitate the molecular characterization of the Herpes simplex Virus types 1 and types 2 genome DNAs, a gene library of cloned restriction frtgments have been produced. The Vero cells were infected with HSV-1 and HSV-2. 48 hours after infection, the infected cells Ivere Iysed, and multinucleated giant cells were observed approximately at seventy-two hours postinfection. The multiplication of HSV-1 and HSV-2 was observed in Vero cells using electromicroscopy. The nucleocapsids in nuclei were obseryed, and the assembled virions were budded out through the vacuole, and the virions were released from the cells. HSV-1 and HSV-2 was analyzed by digestion of their genome DANs with restriction ensymes. HSV-1 and HSV-2 genome DNAs were digested with BarnHI, Bgfl respectively. The BarnHI rlestriction fragments of HSV-1 and HSV-2 genome DNAs were twenty-seven fragments and thair molecular sizes were ranging $0.70{\sim}15.08$, $4.4{\sim}31.0$ tilobases. The BglII restriction fragments of HSV-1 and HSV-2 genome DNAs were sixteen, eighteen fragments and thair molecular sizes were ranging $4.8{\sim}30.0$, $1.2{\sim}25.0$ kilobases. And then BglII restriction frgments were cloned in Escherichia coli(E.coil) using the plasmid vector pBacPAK9.

• Journal of Microbiology and Biotechnology
• /
• 제18권10호
• /
• pp.1630-1633
• /
• 2008

A novel putative toxin-antitoxin segregational stability system named KyAB system was identified in a novel native plasmid pBMB8240 from Bacillus thuringiensis strain YBT-1520, based on sequences homology with other toxin-antitoxin systems, the lethal activity of the KyB putative toxin in Escherichia coli and the stabilizing effect of the kyAB system in Bacillus thuringiensis. Secondarily, the native plasmid pBMB9741 from the same strain was resequenced and the corrected plasmid was named as pBMB7635. Based on sequence homology with the tasAB system and the lethal activity of toxin protein in Escherichia coli, a tasAB-like putative toxin-antitoxin system was identified on pBMB7635.

• 한국양식학회:학술대회논문집
• /
• 한국양식학회 2003년도 추계학술발표대회 논문요약집
• /
• pp.28-28
• /
• 2003

Viable interspecific hybrids between yellowtail flounder (Pleuronectes ferrugineus, Store.) and winter flounder (Pleuronectes americanus, Walbaum) were produced by artificial insemination of yellowtail flounder eggs with winter flounder sperm. However, mean fertilization rate, hatching success and early survival up to 3 weeks post hatch were significantly lower than those of parental pure cross controls (P<0.01). Overall, cytogenetic traits (karyological analysis and estimation of cellular DNA contents using flow cytometry) of hybrid flounder were intermediate between the two parental species. Microsatellite assay was used to distinguish the parental genomes in the hybrids; in most cases, one allele was specific to each of the parents. Morphometrics assessed by body proportions indicated that hybrids generally displayed a morphology intermediate between the maternal and paternal species. Interspecific hybrids exhibited abnormal and retarded gonad development in both sexes based on histological analysis of gonads from adult fish. The sterility of the hybrids presents a significant advantage for their use in aquaculture, as potential escapees would not be capable of reproducing in the wild and contaminating natural stocks.

• Asian-Australasian Journal of Animal Sciences
• /
• 제17권10호
• /
• pp.1366-1368
• /
• 2004

The Mafriwal dairy cattle was developed to meet the demands of the Malaysian dairy Industry. Although there are reports on its production and reproductive performance, there has been no work on its molecular characterization. This study was conducted to characterize the Mafriwal dairy cattle using microsatellite markers. Fifty two microsatellite loci were analysed for forty Mafriwal dairy cows kept at Institut Haiwan Kluang, Malaysia. The study showed two microsatellite loci to be monomorphic. Allele frequencies for the polymorphic loci ranged from 0.01 to 0.31. Genotype frequencies ranged from 0.03 to 0.33. The mean overall heterozygosity was 0.79. All polymorphic microsatellite loci deviated significantly (p<0.01) from Hardy-Weinberg equilibrium. The Mafriwal dairy cattle showed high genetic variability despite being a nucleus herd and artificial insemination being practiced.

• Carbon letters
• /
• 제17권1호
• /
• pp.45-52
• /
• 2016

The attachment of 2-aminobenzamide to carboxylated multi-wall carbon nanotubes (MWCNTs)-COOH was achieved through the formation of amide bonds. Then, the functionalized MWCNTs, MWCNT-amide, were treated by phosphoryl chloride to produce MWCNT-quin. The products were characterized by Fourier transform infrared spectroscopy, Raman spectroscopy, scanning electron microscopy, thermogravimetric analysis, derivative thermogravimetric, steady-state fluorescence spectroscopy, and solubility testing. MWCNT-quin showed photo-electronic properties, which is due to the attachment of the 4-hydroxyquinazoline groups to them as proved by steady-state fluorescence spectroscopy. This suggests intramolecular interactions between the tubes and the attached 4-hydroxyquinazoline. The toxicity of the samples was evaluated in human embryonic kidney HEK293 and human breast cancer SKBR3 cell lines, and the viable cell numbers were measured by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) after the cells were cultured for 24 h. Cellular investigations showed that the modified MWCNTs, particularly MWCNT-quin, have considerably significant toxic impact on SKBR3 as compared to HEK293 at the concentration of 5 µg/mL.

• Asian-Australasian Journal of Animal Sciences
• /
• 제12권1호
• /
• pp.77-83
• /
• 1999

Molecular biological techniques that recently developed, have made it possible to realize some of new attempts in the research field of rumen microbiology. Those are 1) cloning of genes from rumen microorganisms mainly in E. coli, 2) transformation of rumen bacteria and 3) ecological analysis with nonculturing methods. Most of the cloned genes are for polysaccharidase enzymes such as endoglucanase, xylanase, amylase, chitinase and others, and the cloning rendered gene structural analyses by sequencing and also characterization of the translated products through easier purification. Electrotransformation of Butyrivibrio fibrisolvens and Prevotella ruminicola have been made toward the direction for obtaining more fibrolytic, acid-tolerant, depoisoning or essential amino acids-producing rumen bacterium. These primarily required stable and efficient gene transfer systems. Some vectors, constructed from native plasmids of rumen bacteria, are now available for successful gene introduction and expression in those rumen bacterial species. Probing and PCR-based methodologies have also been developed for detecting specific bacterial species and even strains. These are much due to accumulation of rRNA gene sequences of rumen microbes in databases. Although optimized analytical conditions are essential to reliable and reproducible estimation of the targeted microbes, the methods permit long term storage of frozen samples, providing us ease in analytical work as compared with a traditional method based on culturing. Moreover, the methods seem to be promissing for obtaining taxonomic and evolutionary information on all the rumen microbes, whether they are culturable or not.

• 한국환경과학회:학술대회논문집
• /
• 한국환경과학회 2002년도 봄 학술발표대회 발표논문집
• /
• pp.209-212
• /
• 2002

본 연구는 방향족 화합물질 중 페놀폐수에 대한 생물학적 처리를 위해 본 실험실에서 분리한 페놀분해능이 우수한 Rhodococrus sp. EL-GT를 이용하여 catechol 분해 catechol 1,2-dioxygenase 분해활성을 측정하였고, 이것이 ortho-pathway임을 확인할 수 있었다. 또한 다른 연구에서 보고된 Rhodococcus rhodochrous NCIMB 13259 균주의 catechol 1,2 dioxygenase를 기초로한 primer를 이용하여 PCR을 수행하였으며 이 분해 유전자의 cloning실험을 수행 중이다. 이들 실험을 통하여 Rhodococcus sp.의 페놀분해균의 유전적 구조 및 특성을 검토하고 밝혀지는 단백질 정보를 이용하여 방향족 화합물의 분해능이 보다 우수한 균주의 개발을 시도하고자 한다.

• 한국식물병리학회:학술대회논문집
• /
• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
• /
• pp.29-29
• /
• 2003

Incompatible plant-pathogen interactions result in the rapid cell death response known as hypersensitive response (HR) and activation of host defense-related genes. To understand the molecular and cellular mechanism controlling defense response better, several approaches including isolation and characterization of novel genes, promoter analysis of those genes, protein-protein interaction analysis and reverse genetic approach etc. By using the yeast two-hybrid system a clone named Tsipl, Tsil -interacting protein 1, was isolated whose translation product apparently interacted with Tsil, an EREBP/AP2 type DNA binding protein. RNA gel blot analysis showed that the expression of Tsipl was increased by treatment with NaCl, ethylene, salicylic acid, or gibberellic acid. Transient expression analysis using a Tsipl::smGFP fusion gene in Arabidopsis protoplasts indicated that the Tsipl protein was targeted to the outer surface of chloroplasts. The targeted Tsipl::smGFP proteins were diffused to the cytoplasm of protoplasts in the presence of salicylic acid (SA) The PEG-mediated co-transfection analysis showed that Tsipl could interact with Tsil in the nucleus. These results suggest that Tsipl-Tsil interaction might serve to regulate defense-related gene expression. Basically the useful promoters are valuable tools for effective control of gene expression related to various developmental and environmental condition.(중략)

• Molecules and Cells
• /
• 제25권1호
• /
• pp.142-147
• /
• 2008

We recently used degenerate PCR and locus-specific PCR methods to identify the endogenous retroviruses (ERV) in the bovine genome. Using the ovine ERV classification system, the bovine ERVs (BERVs) could be classified into four families. Here, we searched the most recently released bovine genome database with the partial nucleotide sequence of the pro/pol region of the BERV ${\beta}3$ family. This allowed us to obtain and analyze the complete genome of BERV ${\beta}3$. The BERV ${\beta}3$ genome is 7666 nucleotides long and has the typical retroviral organization, namely, 5'-long terminal repeat (LTR)-gag-pro-pol-env-LTR-3'. The deduced open reading frames for gag, pro, pol and env of BERV ${\beta}3$ en- code 507, 271, 879 and 603 amino acids, respectively. BERV ${\beta}3$ showed little amino acid similarity to other betaretroviruses. Phylogenetic analysis showed that it clusters with HERV-K. This is the first report describing the genetic structure and sequence of an entire BERV.

• 대한의생명과학회지
• /
• 제11권1호
• /
• pp.9-13
• /
• 2005

A total of 22 Salmonella enterica serotype Enteritidis (S. Enteritidis) isolates from human and chicken sources were analyzed by pulsed field gel electrophoresis (PFGE) using XbaI restriction enzyme to assess the genetic relationships between strains from different sources. PFGE permitted the resolution of XbaI restriction fragments of the 22 S. Enteritidis into 6 distinct PFGE types (PFT), designated PFT1 to PFT6, and 2 subtypes within PFT2, and allowed to detect between 9 and 10 bands with fragments sizes in the range of $25\~635\;kb$. Four of twelve isolates from human showed an identical PFGE patterns with 2 isolates from chickens. Also, another one isolate from human showed an identical PFGE patterns with other 5 isolates from chickens. Only one isolate from chicken, however, showed a different pattern compared to other PFTs. These results suggested that sporadic human food-poisoning cases infections caused by S. Enteritidis in this study were due to the consumption of contaminated chicken meats and that a clonally highly similar strains exist and spread between human and chicken sources.