• Journal of Aquaculture
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• v.21 no.4
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• pp.259-264
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• 2008

Cytogenetic characteristics of three Hemibarbus species (H. labeo, H. longirostris and H. mylodon) were analyzed based on erythrocyte measurement, flow cytometric estimation of cellular DNA content, and karyological analysis. Average nuclear volumes for H. labeo, H. longirostris and H. mylodon were 22.5, 21.7 and $26.0\;{\mu}m^3$, respectively. The estimated genome sizes of those three species were not significantly different from one another, being recorded as 2.51, 2.33 and 2.35 pg/cell for H. labeo, H. longirostris and H. mylodon, respectively. Modal chromosome numbers of the three species were the same as 2n = 50. However, their karyotypes and fundamental numbers (FN) were different among species; 16M+16SM+18T/A (FN = 82) for H. labeo, 18M+16SM+16T/A (FN = 84) for H. longirostris and 18M+24SM+8T/A (FN = 92) for H. mylodon.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.04a
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• pp.180-180
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• 1996

Previous studies have shown that kainic acid (KA) causes an elevation of hippocampal proenkephalin mRNA level. However, the role of proto-oncogene products, such as c-Fos, c-Jun and Fra proteins in the regulation of KA-induced proenkephalin mRNA increase in the hippocampus has not been well characterized. Thus, in the present study, the effect of cycloheximide (CHX) on KA-induced proenkephalin mRNA and immediate early gene products induction was examined. After pretreating with either vehicle or CHX (20 mg/kg, s.c.) for 30 min, KA (10 mg/kg) was administered s.c. The animals were sacrificed 1,2, or 8 hrs after KA administration. Total RNA and were isolated for Northern blot assay, and proteins were isolated for Western and electrophoretic gel-shift assays. First, we found that CHX inhibited KA-induced proenkephalin mRNA increase without altering intracellular proenkephalin protein level. Secondly, Western blot assays showed that KA increased c-Fos, c-Jun and Fra proteins at 1,2, and 8 hrs and CHX inhibited these immediate early gene products. Finally, electrophoretic gel shift assays revealed that KA increased both AP-1 and ENKCRE-2 DNA binding activities. Furthermore, CHX attenuated KA-induced AP-1 and ENKCRE-2 DNA binding activities. Both AP-1 and ENKCRE-2 DNA binding activities were abolished by cold AP-1 or ENKCRE-2 oligonucleotides, and further reduced by antibodies against c-Fos or c-Jun. Antibody against CREB reduced ENKCRE-2, but not AP-1, DNA binding activity. Our results suggest that on-going protein synthesis is required for elevation of hippocampal proenkephalin mRNA level induced by KA. All c-Fos, c-Jun, and Fra proteins appears to be involved in the regulation of hippocampal proenkephalin mRNA level induced by KA (This study was supported by a grant from KOSEF).

• BMB Reports
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• v.31 no.2
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• pp.149-155
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• 1998

The N199H variant of the EcoRI endonuclease has about twice the catalytic activity of the wild-type. A comparison of their biochemical characteristics, using synthetic oligonucleotides 5'-dAAAACTTAAGAAAAAAAAAAA-3' (KA) and 5'-dTTTTTGAATTCTTTTTTTTTT-3' (KT), helps to define the cleavage reaction pathway of these enzymes. Both EcoRI and EcoRI variant N199H were found to cleave singlestranded KA or KT about three times faster than the double-stranded forms, although the KT oligonucleotide was more susceptible. Using the ssDNA substrate in kinetic analyses, lower $K_m$ values were obtained for the N199H variant than for the wild-type at low (50 mM), as well as high (200 mM), sodium chloride concentrations. This difference between the endonucleases is attributed to a grealter accessibility for tbe substrate by the variant, and also a higher affinity for the DNA backbone. It also appears that the relative activities of the two enzymes, particularly at high ionic strength, are proportional to their populations in the monomeric enzyme form. That is, according to gel filtration data, half of the N199H molecules exist as monomers in 200 mM NaCl, whereas those of the wild-type are mainly dimeric. Consequently, the Asp199 residue of the EcoRI endonuclease may be implicated in the protein-protein interaction leading to dimerization, as well as in coupling to DNA substrates. In summary, it is proposed that active monomeric endonuclease molecules, derived from the dimeric enzyme, recognize and form a complex with a single stranded form of the DNA substrate, which then undergoes nucleophilic substitution and cleavage.

• Journal of Korean Society of Forest Science
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• v.102 no.4
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• pp.537-542
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• 2013

Field performance of somatic plants (somatic embryo derived-plants) of yellow-poplar (Liriodendron tulipifera) produced from somatic embryogenesis was compared with that of seedlings at age 5. In comparison of photosynthetic rate (seedling, $10.67{\mu}mol$ $CO_2m^{-2}s^{-1}$; somatic plant, $9.04{\mu}mol$ $CO_2m^{-2}s^{-1}$), stomatal conductance rate (seedling, 0.2 $H_2Om^{-2}s^{-1}$; somatic plant, 0.166 $H_2Om^{-2}s^{-1}$) and respiration rate (seedling, 1.71 mmol $H_2Om^{-2}s^{-1}$; somatic plant, 1.513 mmol $H_2Om^{-2}s^{-1}$), no significant differences were found between plants. The seedlings were a little higher in comparison of stomatal density (seedling, $23.33/mm^2$; somatic plant, $22.43/mm^2$), length (seedling, $25.83{\mu}m$; somatic plant, $23.46{\mu}m$) and width (seedling, $15.87{\mu}m$; somatic plant, $15.3{\mu}m$). In comparison of DNA content of the leaves using flow cytometry, no differences in ploidy level were found between the seedlings and somatic plants.

• Advances in Traditional Medicine
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• v.9 no.1
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• pp.39-48
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• 2009

We have evaluated the effect of long term treatment of Enicostemma littorale (E. littorale) in type 2 diabetic patients taking pills of aqueous extract of E. littorale regularly as a complimentary medicine for at least 9 months. The effects of E. littorale on glycemic control, lipid profile, cardiac function and DNA damage in these patients were compared with those who had not been regular in taking E. littorale but regular in taking other conventional anti-diabetics. Our data suggest that, E. littorale can maintain normal blood glucose, serum insulin, serum triglycerides levels of type 2 diabetic patients if taken regularly. E. littorale also improves insulin sensitivity, and normalize disturbed lipogram and elevated creatinine levels, thereby produces beneficial effect in preventing cardiovascular complications and may preserve the kidney function. The finding that E. littorale also prevents DNA damage suggest a long term effect in diabetic patients. E. littorale thus can be considered as safe supplementary therapy for a long term and effective management of type 2 diabetic patients.

• The Plant Pathology Journal
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• v.16 no.1
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• pp.9-18
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• 2000

Using differential display techniques, a new acidic pathogenesis-related (PR) protein-1 cDNA (GMPRla) gene was isolated from a cDNA library of soybean (Glycinemax L.Merr, cultivar Jangyup) hypocotyls infected by Phytophthora sojae f. sp. glycines. The 741 bp of fulllength GMPRla clone contains an open reading frame of 525 nucleotides encoding 174 amino acid residues (pI 4.23) with a putative signal peptide of 27 amino acids in the N-terminus. Predicted molecular weight of the protein is 18,767 Da. The deduced amino acid sequence of GMPRla has a high level of identity with PR-1 proteins from Brassica napus, Nicotiana tabacum, and Sambucus nigra. The GMPRla mRNA was more strongly expressed in the incompatible than the compatible interaction. The transcript accumulation was induced in the soybbean hypocotyls by treatment with ethephon or DL-$\beta$-amino-n-butyric acid, but not by wounding. In situ hybridization data showed that GMPRIa mRNAs were usually localized in the vascular bundle of hypocotyl tissues, especially phloem tissue. Differences between compatible and incompatible interactions in the timing of GMPRla mRNA accumulation were remarkable, but the spatial distribution of GMPRla mRNA was similar in both interactions. However, more GMPRla mRNA was accumulated in soybean hypocotyls at 6 and 24 h after inoculation.

• Journal of Microbiology
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• v.37 no.4
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• pp.229-233
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• 1999

The mitochondrial plasmid pMLP1 from a white-rot fungus, Pleurotus ostreatus, is a double-stranded DNA containing 381 bp terminal inverted repeat (TIR) whose 5'-ends are covalently bound by terminal proteins. The plasmid contains two major open reading frames (ORFs), encoding putative DNA and RNA polymerases, and a minor ORF encoding a small, highly basic protein. To identify the DNA binding activity that recognizes the TIR region of pMLP1, gel retardation assays were performed with mitochondrial extracts. A specific protein binding to a region between 123 and 248 nt within TIR was observed. We examined whether the gene product of mORF1 bindes to this region specifically. E. coli cell extract which contains an overproduced mORF1 protein formed a complex specific to the region between 123 and 248 nt. Inclusion of mORF1 protein in the specific complex formed between P. ostreatus mitochondrial extract and TIR was confirmed by a supershift assay using polyclonal antibodies against the mORF1 protein. Our result suggest that the product of mORF1 may function as a terminal region recognition factor (TRF), recognizing an internal region in TIR.

• Korean Journal of Microbiology
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• v.24 no.1
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• pp.18-23
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• 1986

A new type II restriction endonuclease, Bdi I, has been isolated from Brenibacterium divaricatum FERM 5948 by procedures of ammonium sulfate fractionation, DEAE-cellulose chromatography and heparin agarose chromatography. The purified Bdi I restriction endonudlease had the same cleavage patterns of Cla I whose recognition sequence is 5' ATCGAT 3'. From the result that ${\lambda}-Cla$ I DNA frahment could be cloned in pBR 322 digested with Bdi I, it has been proven that Bdi I cuts between T and C(5' AT/CGAT3') within the recognition sequence and produces 5'pCG cohesive end. The optimal temperature for the Bdi I restriction endonuclease activity was $37^{\circ}C$, and optimal salt (NaCl) concentration was 50-100 mM.

• Journal of Life Science
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• v.20 no.8
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• pp.1193-1200
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• 2010

The cDNA cloning and developmental profiles of the mRNA for A. pernyi arylphorin was determined. The complete A. pernyi arylphorin cDNA sequence comprised 2,234 bp (without the poly $A^+$ tail), including an open reading frame of 2,112 bp beginning with a methionine ATG at bp34. The A. pernyi arylphorin contained 704 amino acids which are highly enriched in aromatic amino acids, phenylalanine and tyrosine. The calculated molecular mass of the A. pernyi arylphorin from the ORF was 83,439 Da. The deduced amino acid sequence of A. pernyi arylphorin showed 78, 71, 62 and 64% identity with those of H. cecropia, M. sexta $\alpha$ subunit, M. sexta $\beta$ subunit and B. mori storage protein. In Northern blot analysis, the A. pernyi arylphorin mRNA only in the fat body of the 5th instar larvae was responsible for gene expression of the protein, and the synthetic activity of the mRNA was detected strongly in the early larvae, but not in the middle or late-stage larvae. In addition, a very weak signal in mRNA activity was detected in pupal stages, but this was considered to be inactive mRNA after reviewing the results of the labeling experiment of this protein.

• The Microorganisms and Industry
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• v.11 no.1
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• pp.13-20
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• 1985

특이 단백질(gene II protein)의 정성적 또는 정량적 변화에 의해 DNA 복제에 필요한 replication orgin sequence의 일부분이었단 replication enhancer를 불필요하게 함으로써 minimum replication origin을 core region만으로 국한 되게 함이었다.