• Title/Summary/Keyword: Molecular pattern

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MS2 Labeling of Endogenous Beta-Actin mRNA Does Not Result in Stabilization of Degradation Intermediates

  • Kim, Songhee H.;Vieira, Melissa;Kim, Hye-Jin;Kesawat, Mahipal Singh;Park, Hye Yoon
    • Molecules and Cells
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    • v.42 no.4
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    • pp.356-362
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    • 2019
  • The binding of MS2 bacteriophage coat protein (MCP) to MS2 binding site (MBS) RNA stem-loop sequences has been widely used to label mRNA for live-cell imaging at single-molecule resolution. However, concerns have been raised recently from studies with budding yeast showing aberrant mRNA metabolism following the MS2-GFP labeling. To investigate the degradation pattern of MS2-GFP-labeled mRNA in mammalian cells and tissues, we used Northern blot analysis of ${\beta}$-actin mRNA extracted from the Actb-MBS knock-in and $MBS{\times}MCP$ hybrid mouse models. In the immortalized mouse embryonic cell lines and various organ tissues derived from the mouse models, we found no noticeable accumulation of decay products of ${\beta}$-actin mRNA compared with the wild-type mice. Our results suggest that accumulation of MBS RNA decay fragments does not always happen depending on the mRNA species and the model organisms used.

Cloning and Initial Analysis of Porcine MPDU1 Gene

  • Yang, J.;Yu, M.;Liu, B.;Fan, B.;Zhu, M.;Xiong, T.;Li, Kui
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.9
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    • pp.1237-1241
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    • 2005
  • Mannose-P-dolichol utilization defect 1 (MPDU1) gene is required for utilization of the mannose donor MPD in synthesis of both lipid-linked oligosaccharides (LLOs) and glycosylphosphatidylinositols (GPI) which are important for functions such as protein folding and membrane anchoring. The full length cDNA of the porcine MPDU1 was determined by in silico cloning and rapid amplification of cDNA ends (RACE). The deduced amino acid showed 91% identity to the corresponding human sequence with five predicted transmembrane regions. RT-PCR was performed to detect its expression pattern in five tissues and results showed that it is expressed ubiquitously among the tissues checked. A single nucleotide substitution resulting in the amino acid change (137 Tyr-137 His) was detected within exon 5. Allele frequencies in six pig breeds showed distinctive differences between those Chinese indigenous pigs breeds and European pigs. Using the pig/rodent somatic cell hybrid panel (SCHP), we mapped the porcine MPDU1 gene to SSC12, which is consistent with the comparative mapping result as conservative syntenic groups presented between human chromosome 17 and pig chromosome 12.

DEPDC1 is a novel cell cycle related gene that regulates mitotic progression

  • Mi, Yan;Zhang, Chundong;Bu, Youquan;Zhang, Ying;He, Longxia;Li, Hongxia;Zhu, Huifang;Li, Yi;Lei, Yunlong;Zhu, Jiang
    • BMB Reports
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    • v.48 no.7
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    • pp.413-418
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    • 2015
  • DEPDC1 is a recently identified novel tumor-related gene that is upregulated in several types of cancer and contributes to tumorigenesis. In this study, we have investigated the expression pattern and functional implications of DEPDC1 during cell cycle progression. Expression studies using synchronized cells demonstrated that DEPDC1 is highly expressed in the mitotic phase of the cell cycle. Immunofluorescence assays showed that DEPDC1 is predominantly localized in the nucleus during interphase and is redistributed into the whole cell upon nuclear membrane breakdown in metaphase. Subsequently, siRNA-mediated knockdown of DEPDC1 caused a significant mitotic arrest. Moreover, knockdown of DEPDC1 resulted in remarkable mitotic defects such as abnormal multiple nuclei and multipolar spindle structures accompanied by the upregulation of the A20 gene as well as several cell cycle-related genes such as CCNB1 and CCNB2. Taken together, our current observations strongly suggest that this novel cancerous gene, DEPDC1, plays a pivotal role in the regulation of proper mitotic progression. [BMB Reports 2015; 48(7): 413-418]

Whole Transcriptomic Analysis of Bacillus anthracis during Hydrogen Peroxide Decontamination (과산화수소 제독 과정에서의 탄저균 전사체 분석)

  • Kim, Sang Hoon;Kim, Se Kye;Jung, Kyoung Hwa;Yoon, Sung Nyo;Kim, Yun Ki;Kim, Min Cheol;Ryu, Sam Gon;Lee, Hae Wan;Chai, Young Gyu
    • Journal of the Korea Institute of Military Science and Technology
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    • v.18 no.4
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    • pp.478-483
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    • 2015
  • Decontamination of biological agents utilizes hydrogen peroxide($H_2O_2$) for its effectiveness and safeness. Bacillus anthracis is a major target for $H_2O_2$ decontamination. To assess the effect of $H_2O_2$ on B. anthracis and identify biomarkers for decontamination, whole transcriptomic profiling of $H_2O_2$-treated B. anthracis was performed. Here we identified deregulation in stress response genes, transcription factors and cellular homeostasis genes. We also found that expression of antisense RNAs increased in B. anthracis during decontamination. We postulate that B. anthracis prioritizes survival and adaptation in response to $H_2O_2$ treatment by changing its gene expression pattern.

A Simple Method for Extraction of High Molecular Weight DNA fromPorphyra Tenera (Rhodophyta) Using Diatomaceous Earth

  • Kim, Tae-Hoon;Hwang, Mi-Sook;Song, Ju-Dong;Oh, Min-Hyuk;Moon, Yong-Hwan;Chung, Ik-Kyo;Rhew, Tae-Hyoung;Lee, Choon-Hwan
    • ALGAE
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    • v.21 no.2
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    • pp.261-266
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    • 2006
  • The innate soluble polysaccharides and phenolic compounds of marine macroalgae are serious contaminants which interfere with experimental procedures such as restriction enzyme digestion, polymerase chain reaction (PCR) and other enzymatic reactions using extracted DNA samples. The viscous polysaccharides are co-precipitated with DNA samples by isopropanol or ethanol precipitation in conventional experiment. To overcome the problem, a method for the isolation of high molecular weight DNA from Porphyra tenera is developed with the application of diatomaceous earth column. The isolated DNAs by this method were about 50-100 kb in size and could be digested well with restriction enzymes. The nuclease activity seemed to be minimal, and high reproducibility in the arbitrary primed PCR for RAPD analyses was a distinctive feature. These results suggest that this method is very efficient in isolating nucleic acid from macroalgae including Porphyra.

Biogeographic pattern of four endemic Pyropia from the east coast of Korea, including a new species, Pyropia retorta (Bangiaceae, Rhodophyta)

  • Kim, Sun-Mi;Choi, Han-Gu;Hwang, Mi-Sook;Kim, Hyung-Seop
    • ALGAE
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    • v.33 no.1
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    • pp.55-68
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    • 2018
  • Foliose species of the Bangiaceae (Porphyra s. l.) are very important in Korean fisheries, and their taxonomy and ecophysiology have received much attention because of the potential for developing or improving aquaculture techniques. Although 20 species of foliose Bangiales have been listed from the Korean coast, some of them remain uncertain and need further comparative morphological studies with molecular comparison. In this study, we confirm the distribution of four Pyropia species from the east coast of Korea, Pyropia kinositae, P. moriensis, P. onoi, and P. retorta sp. nov., based on morphology and rbcL sequence data. Although P. onoi was listed in North Korea in old floral works, its occurrence on the east coast of South Korea is first revealed in this study based on molecular data. P. kinositae and P. moriensis, which were originally described from Hokkaido, Japan, are first reported on the east coast of Korea in this study. Pyropia retorta sp. nov. and P. yezonesis share a similar thallus color and narrow spermatangial patches in the upper portion of the frond, and they have a sympatric distribution. However, P. retorta can be distinguished by the curled or twisted thalli and by molecular data. The biogeographic pattern of the two native species, P. kinositae and P. retorta, suggests that the east coast of Korea may have been a place of refugia during the Last Glacial Maximum (LGM), and then recolonized to the northern part of Japan through the restored East Korean Warm Current after the LGM.

Effects of Salinity on Chlorophyll Fluorescence from Porphyra Thalli and Comparison of Species with Different Intertidal Distribution

  • Hong Dang Diem;Kim Tae Hoon;Hwang Mi Sook;Chung Ik Kyo;Lee Choon-Hwan
    • Fisheries and Aquatic Sciences
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    • v.1 no.1
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    • pp.122-128
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    • 1998
  • Characteristic changes in chlorophyll fluorescence from thalli of red alga, Porphyra, under high salt stress and during subsequent recovery were investigated, and the differences in the sensitivity to the stress among four species of Porphyra with different intertidal distributions were compared. By the treatment of NaCl with $9\%$ or higher concentrations, photochemical efficiency (Fv/Fm) decreased in a biphasic pattern: a rapid decrease was observed within 1­3 h and followed by a slow decline. The decrease of Fv/Fm was mainly due to the increase of Fo without significant increases of Fm. When the thalli treated with $15\%$ NaCl for 6 h were returned to natural sea water for recovery, the increase of Fv/Fm also showed a biphasic pattern: a rapid increase of Fv/Fm was observed within 2 h and followed by a slow increase. Differences in the sensitivity to salt stress among the four species could be found during recovery after the treatment of severe salt stress. After the treatment of $20\%$ NaCl for 6 h, Fv/Fm decreased below 0.3 in all of the four Porphyra species, and the species living in upper parts of the intertidal zone (P. suborbiculata and P. pseudolinearis) could recover better compared with the species in lower parts of the intertidal zone (P. seriata and P. yezoensis), during recovery for 24 h. The species collected from the coast of the South Sea seemed to be more tolerant than those in the East Sea.

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Biofunctionality of Peptides Purified from Naturally Fermented Anchovy Sauce (천연 숙성 멸치액젓 Peptide의 생리활성)

  • 박종혁;김상무
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.32 no.7
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    • pp.1120-1125
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    • 2003
  • Bioactive properties of low molecular weight peptides purified from anchovy sauce fermented in underground at 15$\pm$3$^{\circ}C$ for 1, 3, and 5 years, respectively, were investigated. The fermented anchovy sauce for 1 year showed 3 peaks on gel permeation chromatography pattern, while 3 and 5 year fermented anchovy sauce showed 4 and 5 peaks, respectively. The longer fermentation period, the lower molecular weight of peptides on gel permeation chromatography pattern. Antioxidative, antitumor, and ACE inhibitory activities of low molecular weight peptides increased as fermentation period increased. Antioxidative and antitumor activities of peptide peak 3 purified from 3 year fermented anchovy sauce were the highest with 34 and 44 $\mu\textrm{g}$/mL of $IC_{50}$/ values, respectively, while ACE inhibitory activity ($IC_{50}$/, 32 $\mu\textrm{g}$/mL) of peak 3 purified from 1 year fermented was the highest.

Characterization and Expression Pattern of the Partial Myostatin cDNA in Shrimp, Fenneropenaeus chinensis

  • Lee, Sang Beum;Kim, Yong Soo;Yoon, Moongeun;Kim, Su-Kyoung;Jang, In Kwon;Lim, Hyun Jeong;Jin, Hyung-Joo
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.4
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    • pp.224-229
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    • 2007
  • Muscle tissue expresses many muscle-specific genes, including myostatin (also known as GDF8) that is a member of the transforming growth factor-beta superfamily. Myostatin (MSTN) negatively regulates mammalian skeletal muscle growth and development by inhibiting myoblast proliferation. Mice and cattle possessing mutant MSTN alleles display a 'double muscling' phenotype characterized by extreme skeletal muscle hypertrophy and/or hyperplasia. In this study, we first have characterized partial cDNA of a MSTN gene from the muscle tissue in the F. chinensis and examined its expression pattern in various tissues. The partial MSTN gene (GenBank accession number EU 131093) in the F. chinensis was 1134 bp, encoding for 377 amino acids that showed 63-93% amino acid similarity to other vertebrate MSTNs, containing a conserved proteolytic cleavage site (RXRR) and conserved cysteine residues in the C-terminus. Based on a RT-PCR, the MSTN gene was expressed in the all tissues of F. chinensis used in this study.

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Molecular Weight Estimation and Protein Patterns of Panax ginseng, Codonopsis lanceolata and Platycodon grandiflorum by Polyacrylamide gel Electrophoresis (인삼, 사삼 및 도라지의 전기영동에 의한 단백질 패턴과 분자량)

  • Ahn, Sang-Deuk;Chung, Chan-Moon;Kwon, Woo-Saeng;Son, Eung-Ryong;Lee, Yong-Sae;Yoon, Kyeung-Eun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.30 no.4
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    • pp.352-358
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    • 1985
  • This study was carried out to identify the protein patterns and to estimate molecular weights of the ginseng and other plants. Polyacrylamide gradient 2-30% gels for identification of protein patterns and 11% SDS polyacrylamide gels for molecular weight estimation of various plants were used. Proteins extracted with distilled water and phosphate buffer were most clearly separated. Although minor bands were a little different according to kinds of buffer, major bands showed similar pattern. Major protein bands were two bands of 45,000, 66,000 in Panax ginseng, 3 bands of 32,000-39,000 in Codonopsis lanceolata and one double band around 39,000 kd in Platycodon grandiflorum.

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