• Journal of Life Science
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• v.22 no.8
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• pp.1120-1125
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• 2012

Cirsium pendulum plants were collected from Hongcheon, Pyeongchang, Wonju, Yangyang in Kangwondo, Gapyeong in Gyeongkido, and Choongju in Choongcheongbukdo. Cirsium setidens plants were collected from Taebaek in Kangwondo and Bonghwa in Kyeongsangbukdo. Genomic DNA was prepared from those plants and used for the amplification of 18S rDNA, ITS1, 5.8S rDNA, ITS2, and part of 28S rDNA. The PCR products were sequenced, and the sequence was deposited in the GenBank. The comparison of those sequences has revealed that the rDNA sequences are identical for all six C. pendulum plants, but that the ITS1 and ITS2 sequences contain variable nucleotides. The two C. setidens plants had different nucleotides in 18S rDNA, ITS1, and ITS2. The comparison of the DNA sequences of C. pendulum and C. setidens collected in this study with C. pendulum of Hokkaido in Japan and C. japonicum of Anhui in China indicated that the plants of those three species are clearly divided into three distinct groups. The silymarin content of the collected plants was analyzed and turned out to be quite high. Therefore, it has been found that both C. pendulum and C. setidens plants are producing large amounts of silymarin, which has been reported to have various medicinal effects.

• Microbiology and Biotechnology Letters
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• v.51 no.1
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• pp.37-42
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• 2023

Neutral and non-essential amino acid, glutamine (Gln), plays an essential role in supplying nitrogen to all the amino acids and nucleotides in the mammalian body. Gln is also the most important carbon source that provides intermediates for gluconeogenesis and fatty acid synthesis and supplements the tricarboxylic acid cycle in fast-growing cancer cells. Among the known 14 Gln transporter genes, soluted carrier family 1 member 5 (SLC1A5) has been reported to be closely associated with cancer cell growth. Three variants (v1, v2, and v3) have been derived from SLC1A5. Here, we established a heterologous gene expression system for the active form of human SLC1A5 variant-2 (hSLC1A5v2) in Escherichia coli. v2 is the smallest variant that has not yet been studied. Four expression systems were investigated: pBAD, pCold, pET, and pQE. We also addressed the problem of codon usage bias. Although pCold and pET overexpressed hSLC1A5v2 in E. coli, they were functionally inactive. hSLC1A5v2 using the pBAD system was able to catalyze the successful transport of Gln, even if it was not highly expressed. Initial activity of hSLC1A5v2 for [¹⁴C] Gln uptake in E. coli reached up to 6.73 μmole·min^-1·gDW^-1 when the cell was induced with 80 mM L-arabinose. In this study, we demonstrated a heterologous expression system for the human membrane protein, SLC1A5, in E. coli. Our results can be used for the functional comparison of SLC1A5 variants (v1, v2, and v3) in future studies, to facilitae the developement of SLC1A5 inhibitors as effective anticancer drugs.

• Korean Journal of Microbiology
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• v.20 no.4
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• pp.201-209
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• 1982

An enzyme, named GTP cyclohydorlase, that catalizes the hydrolytic removal of carbon No.S of GTP has been partially purified from extracts of Pseudomonas putida (IAM 1506). The enzyme exists in two molecuar weight forms : a high molecular weight form (150,000) and a low molecular weight from (40,000). The high molecular weight form has been purified 25-fold. Some of the properties of the enzyme are as follows : It functions optimally at pH8.0, and at $52^{\circ}C$. The Km value for GTP is $20{\mu}M$. Divalent cations $(Cd^{2+}\;and\;Hg^{2+})$ 2+/) at a concentration of 5mM inhibit completely the enzyme activity. No metal ion including $Mg^{2+}$ is needed for the catalysis. The enzyme is heat labile ; its half at $57^{\circ}C$ is 1.5 min. Of a number of nucleotides tested, only GDP was used to any extent as substrbte in place of GTP. One of the products of the enzyme is determined to be a dihydro-neopterin compound.

• Journal of Microbiology and Biotechnology
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• v.29 no.11
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• pp.1790-1798
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• 2019

Flock House virus (FHV), an insect RNA virus, has a bipartite genome. FHV RNA1 can be packaged in turnip yellow mosaic virus (TYMV) as long as the FHV RNA has a TYMV sequence at the 3'-end. The encapsidated FHV RNA1 has four additional nucleotides at the 5'-end. We investigated whether the recombinant FHV RNA1 could replicate in mammalian cells. To address this issue, we prepared in vitro transcribed FHV RNAs that mimicked the recombinant FHV RNA1, and introduced them into baby hamster kidney (BHK) cells. The result showed that the recombinant FHV RNA1 was capable of replication. An eGFP gene inserted into the frame with B2 gene of the FHV RNA1 was also successfully expressed. We also observed that eGFP expression at the protein level was strong at 28℃ but weak at 30℃. Sequence analysis showed that the 3'-ends of the RNA1 and RNA3 replication products were identical to those of the authentic FHV RNAs. This indicates that FHV replicase correctly recognized an internally-located replication signal. In contrast, the 5'-ends of recombinant FHV RNA1 frequently had deletions, indicating random initiation of (+)-strand synthesis.

• Journal of Plant Biology
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• v.21 no.1_4
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• pp.13-21
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• 1978

The effect of cyclic-AMP on the induction of acid phosphatase activity in barley aleurone layers was examined. Tannic acid was used as a inhibitor. Decursinol and coumarin were also used as a comparison. Maxiumu promotion of the enzyme activity was obtained with 10-5M cyclic-AMP, but this promotion was lower than that of 10-5M GAS induced enzyme activity in incubation medium. The inhibition rate in the addition of tannic acid was shown 17% and 63% at a ratio to GAs (by weight) of 10 : 1, and 58% and 94% at a ratio of 100 : 1 treated with GAs, and cyclic-AMP, respectively. The most potentiation of 10-6M GAS effect was induced by the additiion of suboptimal concentration (10-6M) of cyclic-AMP. Additional GAs and cyclic-AMP were shown the recovery of the enzyme activity inhibited by tannic acid. The combination with cyclic-AMP and theophylline enhanced the enzyme activity, too. Any other nucleotides tested except cyclic-AMP didn't show the action. There were no differences in acid phosphatase isozyme patterns by polyacrylamide disc electrophoresis, in conjunction with the different additions but the size of bands showed great differences. Especially, the 3rd band and the 5th band group were remarkable.

• Korean Journal of Medicinal Crop Science
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• v.13 no.5
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• pp.293-297
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• 2005

A thoredoxin (CTRX) gene was cloned and characterized from a full length cDNA library prepared from taproot of three-year old Codonopsis lanceolata. A CTRX was 666 nucleotides long and has an open reading frame of 372 bp with 124 amino acid residues (pI = 4.92). The deduced amino acid sequence of the CTRX matched to the previously reported plant thioredoxin h genes. The deduced amino acid sequence of CTRX exhibited the similarity of 33-67% among previously registered thioredoxin genes. The expression of CTRX in leaves of Codonopsis lanceolata was increased by wounding and 1 mM $H_2O_2$, but decreased by 0.1 mM cadmium.

• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.275-280
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• 2000

The inulin fructotransferse (depolymerizing) (IFTase, EC 2.4.1.93) gene of Arthrobacter sp. A-6 was cloned and expressed in Escherichia coli and Bacillus subtilis. The IFTase gene consisted of an ORF of 1.311 nucleotides encoding a polypeptide of 436 amino acids containing a signal peptide of 31 amino acids in the N-terminus. The molecular mass of the IFTase based on the nucleotide sequence was calculated to be 46.116 Da. The recombinant E. coli $DH5{\alpha}$ cells expressing the Arthrobacter sp. A-6 IFTase gene produced most of the IFTase intracelularly. In contrast, the recombinant B. subtilis DB 104 carrying the IFTas gene on a B. subtilis-E. Coli expression vector secreted the IFTase into the culture fluid efficiently.

• The Plant Pathology Journal
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• v.21 no.2
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• pp.142-148
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• 2005

Whereas most of isolates of Cucumber mosaic virus(CMV) can induce green mosaic systemic symptoms on zucchini squash, foliar symptoms of a pepper isolate of CMV (Pf-CMV)-infected zucchini squash revealed systemic chlorotic spots. To assess this biological property, infectious full-length cDNA clones of Pf-CMV were constructed using long-template RT-PCR. The complete nucleotide sequences of RNA2 and RNA3 of Pf-CMV were determined from the infectious fulllength cDNA clones, respectively. RNA 2 and RNA3 of Pf-CMV contain 3,070 nucleotides and 2,213 nucleotides, respectively. Overall sequence homology of two RNAs revealed high similarity (90%) between CMV strains, and 60% similarity to those of Tomato aspermy virus and Peanut stunt virus strains. By sequence analysis with known representative strains of CMV, Pf- CMV belongs to a typical member of CMV subgroup IA. The virus has high evolutionary relationship with Fny-CMV, but the pathology of Pf-CMV in zucchini squash was quite different from that of Fny-CMV. The pesudorecombinant virus, F1P2P3, induced chlorotic spot leaf symptom and timing of systemic symptom in squash plants, similar to the plants infected by Pf-CMV. No systemic symptoms were observed when Pf-CMVinoculated cotyledons were removed at 5 days postinoculation (dpi) while Fny-CMV showed systemic symptom at 2 dpi. These results suggest that the pepper isolate of CMV possesses unique pathological properties distinguishable to other isolates of CMVs in zucchini squash.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.18 no.4
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• pp.333-338
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• 1985

In this experiment, the contents of free amino acids, and nucleotides and their related compounds of dried cod products were analysed by amino acid autoanalyzer and HPLC. Proline, histidine, lysine and alanine were dominant amino acids in raw extracts, having $65.4\%$ of total free amino acid contents, but the contents of aspartic acid, serine, isoleucine and phenylalanine were low, and methionine, tyrosine and arginine were detected in trace amount. In free amino acid composition of dried products, abundant amino acids were glycine, histidine, lysine, proline and alanine. Such amino acid as aspartic acid, methionine, tyrosine and phenylalanine were poor. In extracts of storage sample, most free amino acids were increased in both sun dried products and hot air dried products, and glycine, histidine, lysine, proline and alanine were abundant amino acid, such amino acid as methionine, tyrosine and phenylalanine were poor. In raw cod, inosine and IMP were dominant contents which 18.5, $10.0{\mu}mole/g$ on dry base, respectively, but the contents of ATP, ADP, hypoxanthine and AMP were detected less than $3.2{\mu}mole/g$. ATP and ADP were decreased while IMP, inosine and hypoxanthine were increased during drying and storage of cod.

• Korean Journal of Food Science and Technology
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• v.15 no.1
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• pp.1-5
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• 1983

Large quantities of an ascidian, Styela plicata, are harvested annually as the by-product of oyster culture on the south coastal area in Korea. It has been traditionally favored and consumed because of its peculiar taste and flavor. But no study on its taste compounds has been reported. An investigation on the taste compounds of fresh meat and integument of an ascidian has been carried out by a series of analysis of taste compounds including nucleotides, free amino acids, betaine, TMAO, TMA and total creatinine. The most abundant nucleotides in fresh meat were AMP and IMP. Their contents were $1.2\;and\;0.8{\mu}mole/g$, respectively. The contents of nucleotides and their related compounds in integument show similar tendency as in fresh meat but their quantities were very low as compared with those in fresh meat. Seventy two percentage of the total free amino acids in fresh meat was consisted of proline, alanine, glutamic acid, glycine and serine. They occupied also 82% of the total free amino acids in integument, but the amount of free amino acids in integument was about 1/10 of that in fresh meat. The contents of betaine-N and creatinine-N were 6.1 and 2.1% of the extractive-N in fresh meat, respectively. The amounts of TMAO-N and TMA-N were very low contents in fresh meat and trace in integument.