• Title/Summary/Keyword: 임의 돌연변이

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Analysis of Vasopressin Receptor Type 2(AVPR2) Gene in a Pedigree with Congenital Nehrogenic Diabetes Insipidus : Identification of a Family with R202C Mutation in AVPR2 Gene (신성요붕증 가계에서 바소프레신 V2 수용체(AVPR2) 유전자 분석 : AVPR2 유전자 R202C 돌연변이의 발견)

  • Park June-Dong;Kim Ho-Sung;Kim Hee-Joo;Lee Yoon-Kyung;Kwak Young-Ho;Ha Il-Soo;Cheong Hae-Il;Choi Yong;Park Hye-Won
    • Childhood Kidney Diseases
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    • v.3 no.2
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    • pp.209-216
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    • 1999
  • Purpose : Nephrogenic diabetes insipidus (NDI) is a rare X-linked disorder associated with renal tubule resistance to arginine vasopressin (AVP). The hypothesis that the defect underlying NDI might be a dysfunctional renal AVPR2 has recently been proven by the identification of mutations in the AVPR2 gene in NDT patients. To investigate the association of mutations in th AVPR2 gene with NDI, we analyzed the AVPR2 gene located on the X chromosome. Methods : We have analyzed the AVPR2 gene in a kindred with X-linked NDI. The proband and proband's mother were analyzed by polymerase chain reaction-single strand conformational polymorphism(PCR-SSCP) and DNA sequencing of the AVPR2 gene. We also have used restriction enzyme analysis of genomic PCR product to evaluate the AVPR2 gene. Results : C to T transition at codon 202, predictive of an exchange of tryptophan 202 by cysteine(R202C) in the third extracellular domain was identified. This mutation causes a loss of Hae III site within the gene. Conclusion : We found a R202C missense mutation in the AVPR2 gene causing X-linked NDI, and now direct mutational analysis is available for carrier screening and early diagnosis.

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Cloning and Characterization of Dihydroflavonol 4-reductase (DFR) from Matthiola incana R. Br. (Stock(Matthiola incana R. Br.)으로부터 색소유전자의 분리 및 분석)

  • 민병환;김석원;오승철;유장렬
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.5
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    • pp.341-346
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    • 1998
  • In this paper we describe the cloning and expression of the genes encoding the flavonoid-biosynthetic enzyme dihydroflavonol 4-reductase (DFR) in Matthiola incana R. Br. A heterologous cDNA probe from Zea mays was used to isolate full-size DFR cDNA clone from a corolla-specific cDNA library. Comparison of the coding region of this DFR cDNA sequence including the sequences of Zea mays, Anthirrinum majus, Petunia hybrida, Callistephus chinensis, Dianthus caryophyllus and Rosa hybrida reveals a identity higher than 61% at the nucleotide level. The DFR transcript is G/C rich in monocotyledonous plants show a strong codon bias preferring codons with a G or C in the third position. The function of this nucleotide sequences were verified by comparison with amino acid sequences of the amino-terminus and tryptic peptides from purified plant enzyme, by northern blotting with mRNA from wild type and mutant plants and by in vitro expression yielding an enzymatically active reductase. Genomic southern blot analysis showed the presence of one gene for DFR in Matthiola incana. Northern blot analysis of the DFR wild type and mutant lines showed that the lack of DFR activity in the stable acyanic mutant k17b is clearly by a transcriptional block of the DFR gene.

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Salt Stress Responses of an Alanine Aminotransferase Knock-out Mutant of Rice (Oryza sativa L.) (알라닌 아미노기전이효소가 상실된 벼(Oryza sativa L.) 돌연변이체의 고염 스트레스에 대한 반응)

  • Im, Kyoung-Nam;Lee, Chin Bum
    • Journal of Life Science
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    • v.23 no.4
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    • pp.487-494
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    • 2013
  • An AlaAT knock-out mutant (alaat) of rice (Oryza sativa L.) was isolated from T-DNA tagging lines and the genotypes of its progeny were determined with AlaAT1-specific primers. The alaat phenotypes showed decreased growth and grain yield when compared with control plants. The activity of AlaAT1 in the mutant plants was practically undetectable. The responses of alaat plants to growth under salt stress were compared with those of control plants by measuring chlorophyll fluorescence and the activities and mRNA expression of antioxidant enzymes. All abiotic stresses tested (salt, drought, and chilling) caused a similar decrease in chlorophyll fluorescence in both alaat and wild type plants. The activity of peroxidase (POX), an antioxidant enzyme, decreased following salt treatment of alaat plants, while control plant showed an increased activity. The mRNA levels for cAPX (cytosolic ascorbate peroxidase), POX2, and AlaAT were determined by RT-PCR following salt stress. No AlaAT1 mRNA was detected in alaat plants. The POX2 mRNA showed a slightly increased level in the wild type but was not detected in alaat plants, in agreement with the activity assays. The levels of cAPX mRNA were greatly increased in both the wild type and alaat plants. The salt stress effects on rice plant growth are therefore proposed to reflect a loss of function of AlaAT, which alters the activity and synthesis of antioxidant enzymes (especially peroxidases), rather than a direct effect on photosynthesis.

Characterization of a Revertant that Restroes the Export of Ribose-Bnding Potein to the Priplasm in Echerichia coli (리보스 결합 단백질을 페리플라슴으로 수송하는 복귀변이주의 분석)

  • ;;Randall, Linda L.
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.283-290
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    • 1988
  • A spontaneous revertant of mutation rbsB103 that is ribose taxis-positive was characterized. This revertant was found to be export-competent in the export of ribose-binding protein shown by the disappearance of accumulated mutant precursor protein and the export of mature ribose-binding protein to the periplasm. The reversional change was shown to be in the region of risB gene that codes for the amino terminal portion of ribose-binding protein. Analysis by high-performance liquid chromatography of peptide patterns of ribose-binding proteins confirmed the relationship between the wild-type and the revertant proteins as shown for the mutant previously (Iida et al., 1985). When the processing rate of presursor proteins from the wild type and the revertant strain in vivo was compared by pulse-chase experiment, it was found that processing is less efficient than normal in the revertant. Purified mature proteins from both wild-type and revertant were subjected to amino acid sequencing. The results confirmed the amino acid changes deduced from the DNA sequencing and showed that processing of the revertant precursor occured in the correct position even though there are two different amino acids present in the signal sequence.

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Comparison of Effect of Various Types of Soybeans on Mutagenicity and Growth of Human Cancer Cell Lines (콩 종류별 항돌연변이 및 암세포 증식 억제 효과 비교)

  • Lim, Sun-Young
    • Journal of Life Science
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    • v.20 no.10
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    • pp.1532-1537
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    • 2010
  • This study compared the inhibitory effects of methanol extracts from yellow and black soybeans (black soybean, Seomoktae and Seoritae) on mutagenicity using the Ames test and growth of human cancer cells (AGS human gastric adenocarcinoma, HT-29 human colon cancer, Hep 3B hepatocellular carcinoma cells). In the Ames test system using Salmonella typhimurium TA100, aflatoxin $B_1$ ($AFB_1$)-induced mutagenicity was significantly inhibited by treatments with the methanol extracts from either yellow or black soybeans in a dose dependent manner (p<0.05). The methanol extracts from various black soybeans tended to have a greater inhibitory effect compared to those from yellow soybeans. As for N-methyl-N'-nitro-N-nitrosoguamidine (MNNG)-induced mutagenicity, the methanol extracts (5 mg/assay) from black soybean, Seomoktae and Seoritae showed 51%, 61% and 53% inhibitory rates, respectively, indicating that Seomoktae, a type of black soybean, had a stronger antimutagenic activity against mutagens (both $AFB_1$ and MNNG). Methanol extracts from black soybeans showed an inhibitory rate of greater than 50% on the growth of human cancer cells (AGS, HT-29 and Hep 3B) and the inhibition was more effective in the methanol extract from Seomoktae. Our results suggested that the methanol extracts from black soybeans showed stronger inhibitory effects on mutagenicity and growth of cancer cells than those from yellow soybean. It is concluded that intake of black soybean can be recommended for improving health.

A Comparative Study of Physiological Activity and Ingredient Analysis of Glycyrrhiza uralensis Fischer Stems and Leaves Cultivated with Different Wavelength of LED Lights (LED광원에 따른 감초 지상부의 생리활성 및 성분 평가)

  • Bang, Keuk Soo;Chang, Young Nam;Jin, Jong Sik;Park, Sang A;Lim, Jae Soo;Park, Jeong Sub;Kim, Jong Sung;Lee, Jeong Ho
    • Korean Journal of Plant Resources
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    • v.28 no.1
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    • pp.126-134
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    • 2015
  • Glycyrrhiza uralensis Fischer is one of the most commonly used herbs. Recently, the stem and leave of the plant have been interested in physiological activities because the aerial parts have been thrown away. Finding out cultivation method of Glycyrrhiza uralensis Fischer to improve chemical ingredients and biological activities has been tried these days. In this study, different wavelengths of light emitting diode (LED) were used for a cultivation of Glycyrrhiza uralensis Fischer. Antioxidant activities and inhibitory effect on mutagenecity of samples were evaluated. The stem and leave cultivated under blue light (BL-0) showed the strongest antioxidant activities of $3.02{\pm}0.13{\mu}g/ml$ ($EC_{50}$) and $2.18{\pm}0.18{\mu}g/ml$ ($EC_{50}$) in DPPH and ABTS radical scavenging test, respectively. Total phenolic content of BL-0 was $2.93{\pm}0.11g/100g$, the highest value between cultivation conditions. However, antioxidant activities of the stem and leave cultivated under red light were the weakest between samples. All of the stem and leave used in this study showed inhibitory effect on mutagenecity of 1-nitropyrene. BL-0 showed stronger inhibitory effects on mutagenicity of Trp-P-1, Trp-P-2, and AFB1 than samples cultivated under other conditions. Only on mutagenecity of 2-aminoanthracene, the stem and leave cultivated at 1 m apart from red light (RL-1) showed the strongest inhibitory effect. These results indicate that blue LED might be the most effective condition for improvement of physiological activities for the aerial parts of Glycyrrhiza uralensis Fischer in cultivation. The components were identified with GC/MS. Cytidine was detected only in RL-1 at 25 min of retention time and 2-bromotrimethylene glycol was detected only in BL-0 at 37 min.

Overproduction and Purification of Ribose-Binding Proteins from the Wild-Type Mutant and Revertant Strains in Escherichia coli (리보스 결합단백질의 대량생산을 위한 야생형 수송결합변이, 복귀변이 유전자의 클로닝과 이들 단백질의 순수정제)

  • ;Randall Linda L.
    • Korean Journal of Microbiology
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    • v.26 no.4
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    • pp.291-297
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    • 1988
  • Three alleles of rbsB gene, rbsB, rbsB103, and rbsB106 from the wild type, the mutant and the revertant strain, respectively, were cloned for overproduction of proteins under the control of lambda $P_{L}$ promoter. Five different species of precursor and mature ribose-binding proteins were purified to homogeneity using DEAE-Sephadex column chromatography, osmotic shock pocedure, CM-Sephadex column chromatography, and Chromatofocusing column chromaography. pI of the precursor proteins and mature proteins were determined and found to be pH 8.0 and 7.5, respectively. The purified proteins were subjected to amino acid sequencing. The results confirmed the amino acid changes deduced from the DNA sequencing.

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Isolation of SYP61/OSMl that is Required for Salt Tolerance in Arabidopsis by T-DNA Tagging (애기장대에서 고염 스트레스 내성에 관여하는 OSM1/SYP61 유전자의 동정)

  • Kim, Ji-Yeon;Baek, Dong-Won;Lee, Hyo-Jung;Shin, Dong-Jin;Lee, Ji-Young;Choi, Won-Kyun;Kim, Dong-Giun;Chung, Woo-Sik;Kwak, Sang-Soo;Yun, Dae-Jin
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.11-18
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    • 2006
  • Salt stress is one of major environmental factors influencing plant growth and development. To identify salt tolerance determinants in higher plants, a large-scale screen was conducted with a bialaphos marker-based T-DNA insertional collection of Arabidopsis ecotype C24 mutants. One line for salt stress-sensitive mutant (referred to as ssm1) exhibited increased sensitivity to both ionic (NaCl) and nonionic (mannitol) osmotic stress in a root growth assay. This result suggests that ssm1 mutant is involved in ion homeostasis and osmotic compensation in plant. Molecular cloning of the genomic DNA flanking T-DNA insert of ssm1 mutant was achieved by mutant genomic DNA library screening. T-DNA insertion appeared in the first exon of an open reading frame on F3M18.7, which is the same as AtSYP61. SSM1 is SYP61/OSM1 that is a member of the SNARE superfamily of proteins required for vesicular/target membrane fusions and factor related to abiotic stress.

Antagonistic Relationships Between Development of Sporangiophores and Phototropism Depending on Monochromatic Light Wavelengths in Phycomyces blakesleeanus (털곰팡이(Phycomyces blakesleeanus)의 생장기간 중 파장별 단색광 조사에 따른 굴광성 반응과 포자낭병 발생간의 길항관계)

  • Lee, Bum-Kyu;Park, Ae-Ri;Kim, Hye-Jeong;Shin, Dong-Ho;Choi, Kwan-Sam
    • Korean Journal of Agricultural Science
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    • v.36 no.1
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    • pp.27-40
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    • 2009
  • Six strains of Phycomyces blakesleeanus (DEL, A56, C9, C47, C111, and G5) were cultured in continuous light condition for 6 days. The increase in fresh and dry weight after the culture period was similar in all strains. However, the increase in fresh weight was sustained throughout the culture period in all strains, whereas the increase in dry weight was prominent only during the first 3 days of culture. The development and phototropism of macrophores was observed after 3 days of culture, These results suggest that the development and phototropism is ascribed to the increase in turgor pressure in sporangiophores where water entered. A56 and G5 showed a higher sensitivity to light than C9 and C47, whereas C111 showed no photosensitivity. Antagonistic relations were observed between photosensitivity and development of microphores, suggesting that the development and phototropism of macrophores is regulated by a common recognition factor (photoreceptor).

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Antimutagenic and Anticancer Effects of Salted Mackerel with Various Kinds of Salts (소금의 종류에 따른 염장 고등어의 항돌연변이효과 및 암세포 성장억제효과)

  • KONG Chang-Suk;BAK Soon-Sun;JUNG Keun-Ok;KIL Jeung-Ha;LIM Sun-Young;PARK Kun-Young
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.38 no.5
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    • pp.281-285
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    • 2005
  • Raw meat of mackerel (Scomber japonicus) was salted by refined, sun-dried, bamboo, and KC1-added bamboo salts. Antimutagenic activity on N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in Ames test and growth inhibitory effects of AGS human gastric and HT-29 human colon adenocarcinoma cells were investigated using methanol extracts of the salted mackerels. Mackerel salted sun-dried, bamboo, and KC1-added bamboo salts used increased the antimutagenic activities against MNNG, however, the sample treated with refined salt reduced the antimutagenic activity. Inhibitory effects of the salted-mackerels on the growth of human cancer cells were increased as dose dependent pattern. Mackerel salted with refined salt activated the growth of AGS human gastric adenocarcinoma cells, but mackerel salted with sun-dried, bamboo, and KC1-added bamboo salts kept or increased anticancer effect compared to the raw mackerel. Mackerel salted with KC1-added bamboo salt led to the highest antimutagenic and anticancer activities. These results suggest that antimutagenic and anticancer effects of mackerel during manufacturing of the salted-mackerel could be enhanced by using different kind of salts such as bamboo, or KC1-added bamboo salts.