• Title/Summary/Keyword: P gene

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Characterization of Transgenic Tall Fescue Plants Expressing Two Antioxidant Genes in Response to Environmental Stresses (두 가지 항산화유전자를 동시에 발현시킨 형질전환 톨 페스큐 식물체의 환경스트레스에 대한 내성 특성 해명)

  • Lee, Sang-Hoon;Lee, Ki-Won;Kim, Ki-Yong;Choi, Gi-Jun;Seo, Sung;Kwak, Sang-Soo;Kwon, Suk-Yoon;Yun, Dae-Jin;Lee, Byung-Hyun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.27 no.2
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    • pp.109-116
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    • 2007
  • Environmental stress is the major limiting factor in plant productivity. As an effort to solve the global food and environmental problems using the plant biotechnology, we have developed transgenic tall fescue (Festuca arundinacea Schreb.) plants via Agrobacterium-mediated gene transfer method. To develop transgenic tall fescue plants with enhanced tolerance to the environmental stresses, both CuZn superoxide dismutase (CuZnSOD) and ascorbate peroxidase (APX) genes were incorporated in a pIG121 binary vector and the both of the genes were controlled separately by an oxidative stress-inducible sweet potato peroxidase 2 (SWPA2) premoter expressed in chloroplasts. Leaf discs of transgenic plants showed 10-30% less damage compared to the wild-type when they exposed to a wide range of environmental stresses including methyl viologen (MV), $H_2O_2$ and heavy metals. In addition, when $200{\mu}M$ MV was sprayed onto the whole plants, transgenic plants showed a significant reduction of visible damage compared to wild-type plants that were almost damaged. These results suggest that over expression of CuZnSOD and APX genes in transgenic plants might be a useful strategy to protect the crops against a wide range of environmental stresses.

Expression of MicroRNA-221 in Korean Patients with Multiple Myeloma (한국인의 다발성골수종 환자에서 MicroRNA-221의 발현)

  • Choi, Woo-Soon
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.2
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    • pp.197-204
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    • 2018
  • Multiple myeloma (MM) is the leading cause of death among hematologic neoplasms. Recently, microRNA has been reported to be useful in the diagnosis of multiple myeloma. This study examined whether miR-221 could be used as a diagnostic marker for multiple myeloma. The study was performed on 20 patients with multiple myeloma without any other hematological diseases. MicroRNA extraction was performed using formalin-fixed paraffin-embedded (FFPE) tissues obtained from the bone marrow of patients with multiple myeloma. miR-15a, miR-16, miR-21, miR-181a, and miR-221 were selected as the microRNA target genes for multiple myeloma. The significance of microRNA was based on a fold change of <1.5. To quantify the fold changes, data normalized to the human gene, SNORD43, were used as the values of the patient group. Fold change values greater than 1.5 were defined as "overexpression", whereas values less than -1.5 were defined as "underexpression". Of note, 65.0% (13/20) of samples showed significant "overexpression" in the levels of miR-221 expression and plasma cells with a group of more and less than 30% in MM patients did not show any significance of plasma cell (P<0.05). The results of other studies showing a correlation between the expression of miR-221 and MM in Caucasians were confirmed. These results suggest that miR-221 may be a useful indicator for diagnosing patients with MM. In conclusion, miR-221 is useful in the diagnosis and determining the prognosis of multiple myeloma in Koreans.

Identification and Chararterization of Stationary-phase Specific Cytosolic Protein in Salmonella typhimurium (정지기 Salmonella typhimurium 세포에서 특이적으로 발현되는 세포질 단백질의 동정 및 발현조절에 대한 연구)

  • Yoo, Ah-Young;Kim, Young-Hee;Yu, Jong-Earn;Kim, Sam-Woong;Baik, Hyung-Suk;Kang, Ho-Young
    • Journal of Life Science
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    • v.17 no.2 s.82
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    • pp.298-304
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    • 2007
  • Salmonella is facultative intracellular pathogen that can survive and replicate in macrophages even though these cells are equipped with a plethora of anti-microbial mechanisms. To survive in this hostile intracellular environment, Salmonella has evolved numerous defense mechartisms. An approximately 20 kDa protein was detected as a stationary-phase specific protein band in cytosolic fraction. It was identified as a DNA binding protein in stationary phase (Dps) by analysis of MALDI-TOF assay. It has been known that Dps, the protein produced in the stationary phase of bacteria, allows DNA to form chromatin by binding to DNA nonspecifically and protects DNA from reactive oxidative species (ROS). For further study, Dps specific polyclonal antibodies were generated by injection of purified Dps protein into rabbit. To examine the Finfluence of several regulatory proteins in the expression dps gene, Dps protein level in various S. typhimurium mutants defecting regulatory proteins were investigated by Westernblot using Dps specific polyclonal antibodies.

Expression and Production of Human Granulocyte Colony Stimulating Factor (G-CSF) in Silkworm Cell Line (누에세포를 이용한 인간 G-CSF의 발현 및 생산)

  • Park, Jeong-Hae;Jang, Ho-Jung;Kang, Seok-Woo;Goo, Tae-Won;Chung, Kyung-Tae
    • Journal of Life Science
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    • v.20 no.11
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    • pp.1577-1581
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    • 2010
  • Granulocyte colony stimulating factor (G-CSF) is a hematopoietic cytokine that stimulates bone marrow cells to proliferate and differentiate into granulocytes. G-CSF is approved and used for therapeutic purposes. The endoplasmic reticulum (ER) signal peptide of hG-CSF was replaced with silkworm-specific signal peptides to express and efficiently secrete recombinant hG-CSF by silkworm cells. Plasmids that contain cDNAs for hG-CSF and hG-CSF fused with silkworm- specific signal peptides of prophenoloxidase activating enzyme (PPAE), protein disulfide isomerase (PDI), and bombyxin (BX) were constructed. The G-CSF protein was expressed in insect cell line BM5 and was detected by western blot analysis. The cells transfected with plasmids containing rhG-CSF genes with silkworm-specific signal sequences released mature rhG-CSF protein more efficiently than the cells transfected with pG-CSF, the plasmid containing human G-CSF gene, including its own signal sequence. The production of hG-CSF reached maximal level at four days post-transfection and remained at a high level until 7 days post-transfection. These data demonstrate that the modification of the human G-CSF mimic to insect proteins synthesized in ER greatly improves the production of the protein.

Identification of Methicillin-Resistant Staphylococcus aureus by Polymerase Chain Reaction (중합효소 연쇄반응을 이용한 메치실린 내성균주의 동정)

  • Park, In-Cheol;Kim, Gwang-Su;Park, Myeong-Jin;Lee, Seung-Hun;Hong, Seok-Il;Choe, Tae-Bu
    • KSBB Journal
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    • v.14 no.4
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    • pp.460-464
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    • 1999
  • Methicillin-resistant Staphyloccus aureus (MRSA) has been known to be resistant to many kinds of antibiotics and causes a problem of nosnocomial infection since the third generation of cephalosporines has been introduced in the 1980s. As antibiotic sensitivity tests which have been routinely used to detect MRSA in the laboratory depend on the culture conditions such as, pH, temperature, and time, etc., it is difficult to decide in the case of borderline- or low-level of MRSA. Therefore it would be necessary to develope a new method based on the molecular biological technique to overcome these problems. In this study, we extracted DNA from S. aureus and performed polymerase chain reaction (PCR) to amplify mec A gene, encoding penicillin-binding protein 2' (PBP-2'), which is known to confer bacteria resistance to the bacteriostatic action of methicillin. The results were compares with those of minimal inhibitory concentration (MIC) test. When MIC test with oxacillin was performed on the 120 isolates of S. aureus from each patient's specimens, 64 of them were MRSA and 56 of them were methicillin-sensitive Staphylococcus aureus (MSSA). In pus specimen, more precisely, 61.9% (26/42) of MRSA was detected, and 44.2% (19/43), 60% (9/15) and 50% (10/20) of MRSA were detected in sputum, body fluid, and other specimen respectively. When 40 isolates of MRSA and MSSA were tested by PCR method and compares with the results of MIC method, different results were obtained from 1 isolate of MRSA (2.5%) and in 2 isolates of MSSA (5%) suggesting that PCR method should be performed at the same time for more accurate clinical test of MRSA.

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Mitochondrial Genetic Diversity and Phylogenetic Relationships of Siberian Flying Squirrel(Pteromys volans) Populations

  • Lee, Mu-Yeong;Park, Sun-Kyung;Hong, Yoon-Jee;Kim, Young-Jun;Voloshina, Inna;Myslenkov, Alexander;Saveljev, Alexander P.;Choi, Tae-Young;Piao, Ren-Zhu;An, Jung-Hwa;Lee, Mun-Han;Lee, Hang;Min, Mi-Sook
    • Animal cells and systems
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    • v.12 no.4
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    • pp.269-277
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    • 2008
  • Siberian flying squirrel, an endangered species in South Korea, is distributed through major mountain regions of South Korea. The number of Siberian flying squirrel(Pteromys volans) in South Korea has decreased and their habitats are fragmented and isolated because of anthropogenic activities. So far no molecular genetic data has, however, been available for their conservation and management. To obtain better information concerning genetic diversity and phylogenetic relationships of the Siberian flying squirrel in South Korea, we examined 14 individuals from South Korea, 7 individuals from Russia, and 5 individuals from northeastern China along with previously published 29 haplotypes for 1,140 bp of the mtDNA cytochrome b gene. The 14 new individuals from South Korea had 7 haplotypes which were not observed in the regions of Russia and Hokkaido. The level of genetic diversity(0.616%) in the South Korean population was lower than that in eastern Russia(0.950%). The geographical distribution of mtDNA haplotypes and reduced median network confirmed that there are three major lineages of Siberian flying squirrel, occupying; Far Eastern, northern Eurasia, and the island of Hokkaido. The South Korean population only slightly distinct from the Eurasia, and eastern Russian population, and is part of the lineage Far Eastern. Based on these, we suggest that the South Korean population could be considered to belong to one partial ESU(Far Eastern) of three partial ESUs but a different management unit. However, the conservation priorities should be reconfirmed by nuclear genetic marker and ecological data.

Immunohistochemical studies on the relationship between pineal body and superior cervical ganglia of the Korean native goat (한국재래산양 송과체와 앞쪽목신경절의 관계규명을 위한 면역조직화학적 연구)

  • Lee, Heungshik S.;Lee, In-Se;Song, Seung-hoon;Yoon, Sung-tae;Hwang, In-koo;Lee, Choong-hyun
    • Korean Journal of Veterinary Research
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    • v.40 no.2
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    • pp.197-211
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    • 2000
  • The pineal body have been known to be affected by superior cervical ganglia, and most of its nerve fibers containing peptidergic neurotransmitters have been considered to be originated from this ganglia. To confirm this relationships, some peptidergic neurotransmitters were identified in both of pineal body and superior cervical ganglia of the Korean native goat, which were divided into two group; breeding season and non-breeding season. The localizations of two catecholamine-synthesizing enzymes; tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH), were investigated by immunohistochemistry in the superior cervical ganglia and the pineal body of adult Korean native goats. Substance P (SP), calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY) and galanin (GAL) were also identified in these organs by immunohistochemical and double immunofluorescent methods. In superior cervical ganglia, immunoreactivities for TH and DBH were confirmed in the same ganglion cells. The immunoreactivites for SP, VIP(only in male), NPY and GAL were identified in both of ganglion cell bodies and nerve fibers in the ganglia. CGRP immunoreactivity, however, was observed only in nerve fibers. Most NPY- and VIP-immunoreactive(IR) ganglion cells also contained TH. SP and TH were colocalized in the cell bodies, but not in the nerve fibers. TH immunoreactivity was shown in almost all of ganglion cells in the superior cervical ganglia. The immunoreactivity for NPY had some seasonal variation and was stronger in breeding season than in non-breeding season. In pineal body, lots of TH-IR fibers were observed throughout the parenchyma including the pineal stalk and most of them also contained DBH. SP- and NPY-IR fibers were also immunostained with TH or DBH. But a few SP- and NPY-IR fibers were not colocalized with TH or DBH. Exceptionally, a bipolar neuron-like cell was observed to be immunostained with NPY in the pineal body. A few CGRP and GAL-IR fibers were observed, while VIP-IR fibers were not present. It is concluded that most TH- and DBH-IR fibers as well as the peptidergic immunoreactive fibers of the pineal body might be originated from the superior cervical ganglia. Some peptidergic immunoreactive fibers, however, might be come from other regions of brain. We also suggest that NPY in pineal body plays a important role for pineal function. The seasonal variation of NPY immunoreactivity indicates that the synthesis and use of NPY may be different between in breeding and non-breeding seasons.

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Biological Characteristics and Tissue Structure of a Crustose Coralline Lithophyllum Alga (해조류 무절산호조 혹돌잎의 생물학적 특성 및 조직구조)

  • Kang, Ji-Young;Benliro, Ianthe Marie P.;Lee, Ik-Joon;Choi, Ji-Young;Joo, Jin;Choi, Yoo Seong;Hwang, Dong Soo;Hong, Yong-Ki
    • Journal of Life Science
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    • v.23 no.3
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    • pp.341-346
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    • 2013
  • The disappearance of seaweed flora in some rocky areas, which is known as algal whitening, barren ground, coralline flats, or deforested areas, is associated with some species of coralline algae. To determine the biological characteristics of a representative species of crustose coralline alga, the 18S rDNA gene was sequenced to identify the genus Lithophyllum. According to its morphological and distributional characteristics, it was deduced to be L. yessoense. Viability was measured using triphenyl tetrazolium chloride and showed high viability from December to February. Culture conditions of $16^{\circ}C$, a 16 hr light, 8 hr dark cycle, and 30 ${\mu}E/m^2/s$ light intensity were optimal for maintaining the viability of the alga for up to five days. Included in the fatty acids was 9.7% ${\omega}$-3 eicosapentaenoic acid. An electron microscopy scan of the surface structure revealed round craters about 3.6 ${\mu}m$ in diameter, which were covered with rough, irregular, and angular polygon-shaped structures about 1.0 to 3.7 ${\mu}m$ in size. Based on the composition and structure found in our study, biomimetic coralline alga might become an environmentally friendly antifouling material against the attachment of soft foulants.

Isolation and Characterization of Plant-Derived Lactic Acid Bacteria as Potential Probiotic (잠재적 생균제로서 식물 젖산균의 분리 및 특성)

  • Kim, Jeong-Do;Park, Sung-Bo;Lee, Na-Ri;Jeong, Jin-Ha;Lee, Hee-Seob;Hwang, Dae-Youn;Lee, Jong-Sup;Jeong, Seong-Yun;Son, Hong-Joo
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.308-312
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    • 2011
  • Plant lactic acid bacteria were isolated from plant-associated fermentative foods and crops, and their probiotic properties were investigated. Isolates K27 and O2 were isolated from Kimchi and onion, and identified as Lactobacillus plantarum on the basis of 16S rRNA gene analysis. The two strains were highly resistant to acid (an MRS broth at pH 2.5), where the survival rates of L. plantarum K27 and L. plantarum O2 were 90.2% and 97.3%, respectively. L. plantarum K27 and L. plantarum O2 also showed high bile resistance to 0.5% oxgall, with a more than 70% survival rate. They showed an inhibitory effect against pathogenic strains of Escherichia coli KCCM 40880 and Pseudomonas aeruginosa ATCC 10145. The antibacterial effect of the two strains was probably due to the presence of lactic acid. ACE inhibitory activities of the two strains ranged from 72.8% to 80.6% in MRS broth. Notably, the two strains showed high ACE inhibitory activity (89.2~98.2%) in MRS broth containing 10% skim milk. Antioxidant activity was tested by DPPH radical scavenging activity, with antioxidant activities of the strains being in the range of 56.8~61.5%. The results obtained in this study suggest that L. plantarum K27 and L. plantarum O2 may be potential probiotic starter cultures with applications with fermentative products.

Antibacterial Activity and Probiotic Potential of Lactobacillus plantarum HKN01: A New Insight into the Morphological Changes of Antibacterial Compound-Treated Escherichia coli by Electron Microscopy

  • Sharafi, Hakimeh;Maleki, Hadi;Ahmadian, Gholamreza;Zahiri, Hossein Shahbani;Sajedinejad, Neda;Houshmand, Behzad;Vali, Hojatollah;Noghabi, Kambiz Akbari
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.225-236
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    • 2013
  • Among several bacteria examined, an antibacterial-producing Lactobacillus strain with probiotic characteristics was selected and identified based on 16S rRNA gene sequencing. Subsequent purification and mode of action of the antibacterial compounds on target cells including E. coli were investigated. Maximum production of the antibacterial compound was recorded at 18 h incubation at $30^{\circ}C$. Interestingly, antibacterial activity remained unchanged after heating at $121^{\circ}C$ for 45 min, 24 h storage in temperature range of $70^{\circ}C$ to room temperature, and 15 min exposure to UV light, and it was stable in the pH of range 2-10. The active compounds were inactivated by proteolytic enzymes, indicating their proteinaceous nature, and, therefore, referred to as bacteriocin-like inhibitory substances. Isolation and partial purification of the effective agent was done by performing ammonium sulfate precipitation and gel filtration chromatography. The molecular mass of the GFC-purified active compound (~3 kDa) was determined by Tris-Tricine SDS-PAGE. To predict the mechanisms of action, transmission electron microscopy (TEM) analysis of ultrathin sections of E. coli before and after antibacterial treatment was carried out. TEM analysis of antibacterial compounds-treated E. coli demonstrated that the completely altered bacteria appear much darker compared with the less altered bacteria, suggesting a change in the cytoplasmic composition. There were also some membrane-bound convoluted structures visible within the completely altered bacteria, which could be attributed to the response of the E. coli to the treatment with the antibacterial compound. According to the in vivo experiments oral administration of L. plantarum HKN01 resulted in recovery of infected BALB/c mice with Salmonella enterica ser. Typhimurium.