• Title/Summary/Keyword: Molecular Characteristics

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Molecular Cloning of Insulin-like Growth Factor-I (IGF-I) and IGF-II Genes of Marine Medaka (Oryzias dancena) and Their Expression in Response to Abrupt Transfer from Freshwater to Seawater

  • Kang, Yue-Jai;Kim, Ki-Hong
    • Fisheries and Aquatic Sciences
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    • v.13 no.3
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    • pp.224-230
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    • 2010
  • Growth hormone (GH) is known as one of the main osmoregulators in euryhaline teleosts during seawater (SW) adaptation. Many of the physiological actions of GH are mediated through insulin-like growth factor-I (IGF-I), and the GH/IGF-I axis is associated with osmoregulation of fish during SW acclimation. However, little information is available on the response of fish IGF-II to hyperosmotic stress. Here we present the first cloned IGF-I and IGF-II cDNAs of marine medaka, Oryzias dancena, and an analysis of the molecular characteristics of the genes. The marine medaka IGF-I cDNA is 1,340 bp long with a 257-bp 5' untranslated region (UTR), a 528 bp 3' UTR, and a 555-bp open reading frame (ORF) encoding a propeptide of 184 amino acid (aa) residues. The full-length marine medaka IGF-II cDNA consists of a 639 bp ORF encoding 212 aa, a 109 bp 5' UTR, and a 416 bp 3' UTR. Homology comparison of the deduced aa sequences with other IGF-Is and IGF-IIs showed that these genes in marine medaka shared high structural homology with orthologs from other teleost as well as mammalian species, suggesting high conservation of IGFs throughout vertebrates. The IGF-I mRNA level increased following transfer of marine medaka from freshwater (FW) to SW, and the expression level was higher than that of the control group, which was maintained in FW. This significantly elevated IGF-I level was maintained throughout the experiment (14 days), suggesting that in marine medaka, IGF-I is deeply involved in the adaptation to abrupt salinity change. In contrast to IGF-I, the increased level of marine medaka IGF-II mRNA was only maintained for a short period, and quickly returned a level similar to that of the control group, suggesting that marine medaka IGF-II might be a gene that responds to acute stress or one that produces a supplemental protein to assist with the osmoregulatory function of IGF-I during an early phase of salinity change.

Morphological and Molecular Characterization of Heterodera schachtii and the Newly Recorded Cyst Nematode, H. trifolii Associated with Chinese Cabbage in Korea

  • Mwamula, Abraham Okki;Ko, Hyoung-Rai;Kim, Youngjoon;Kim, Young Ho;Lee, Jae-Kook;Lee, Dong Woon
    • The Plant Pathology Journal
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    • v.34 no.4
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    • pp.297-307
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    • 2018
  • The sugar beet cyst nematode, Heterodera schachtii is a well known pathogen on Chinese cabbage in the highland fields of Korea. However, a race of cyst forming nematode with close morphological resemblance to H. trifolii was recently isolated from the same Chinese cabbage fields. Morphological species differentiation between the two cyst nematodes is challenging, with only minor differences between them. Thus, this study described the newly intercepted H. trifolii population, and reviewed morphological and molecular characteristics conceivably essential in differentiating the two nematode species. A comparison of morphometric measurements of both infective juveniles and vulval cones of cysts showed significant differences between the two cyst nematodes. When total RNA and genomic DNA were extracted from a mixed field population, COI genes and ITS regions were clearly amplified with primers of the two Heterodera species, suggesting that Heterodera population collected from the Chinese cabbage field consisted of a mixture of two species. COI and ITS of H. trifolii were predominantly amplified from nucleotides prepared from H. trifolii monoxenic population whereas those of H. schachtii were strongly detected in H. schachtii monoxenic cultures. Thus, this study confirms the coexistence of the two species in some Chinese cabbage fields; and the presence of H. trifolii in Korea is reported here for the first time.

First record of a marine microalgal species, Micractinium singularis (Trebouxiophyceae) isolated from Janghang Harbor, Korea

  • Jo, Seung-Woo;Kang, Nam Seon;Chae, Hyunsik;Lee, Jung A;Kim, Kyeong Mi;Yoon, Moongeun;Hong, Ji Won;Yoon, Ho-Sung
    • Korean Journal of Environmental Biology
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    • v.38 no.1
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    • pp.61-70
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    • 2020
  • A eukaryotic microalga was isolated from seawater in Janghang Harbor, Korea and its morphological, molecular, and physiological characteristics were investigated. Due to its simple morphology, no distinctive characters were found by morphological observation, such as light microscope or scanning/transmission electron microscopy (S/TEM). However, molecular phylogenetic evidence inferred from the concatenated small subunit (SSU) 18S rRNA and internal transcribed spacer (ITS) sequence data indicated that the isolate belonged to the newly described Micractinium singularis. Furthermore, it was clustered with Antarctic Micractinium strains and it also showed a psychrotolerant property, surviving at temperatures as low as 5℃. However, its optimal growth temperatures range from 15℃ to 25℃, indicating that this microalga is a mesophile. Additionally, gas chromatography-mass spectrometry (GC/MS) analysis showed that the isolate was rich in nutritionally important omega-3 polyunsaturated fatty acid, and high-performance liquid chromatography analysis (HPLC) revealed that the high-value antioxidant lutein was biosynthesized as an accessory pigment by this microalga, with glucose as the major monosaccharide. Therefore, in this study, a Korean marine M. singularis species was discovered, characterized, and described. It was subsequently added to the national culture collections.

A Study on the isolation and characteristics of fibrinolysis-related enzymes from Holotrichia extract (제조의 혈전(血栓) 용해(溶解) 효소(酵素) 분리(分離) 및 그 특성(特性)에 관한 연구(硏究))

  • Hong, Shi-Nae;Jeong, Ji-Cheon;Kim, Cheol-Ho
    • The Journal of Internal Korean Medicine
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    • v.20 no.1
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    • pp.198-209
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    • 1999
  • A thrombus is a mass formed from the constituents of the blood within the vessels or the heart during life. The process of its formation is known as thrombosis, It has been generally accepted that Holotrichia is an useful medicine for thrombosis. The rate of fibrinolysis of Holotrichia extracts increase as incubation times. Especially 52 days, the effect on the extracts has an maximum increased fibrinolytic activity, Heat-and-pH-stability of the extract on fibrinolysis is relative to temperature, At $37^{\circ}C$, it has activating effect between pH 4 and pH 12. At higher temperature, especially $80^{\circ}C$, an excessive increase in temperature has a deactivating effect on the extracts. Optimal pH of the extract on fibrinolysis is between pH 7.0 and pH 8.5, it is effective within a relatively broad pH range. In experiments of various inhibitors of Holotrichia extracts fibrinolytic activity, there are strong inhibitive effect on SBTI and Aprotinin, and a few inhibitive effect on DFP and t-AMCHA, no effect on PSMB and TLCK. Holotrichia extracts mixing with fibrinogen are observed Electron microscopy. it shows partially erosive-shaped fibrinolytic activity. In a SDS-PAGE of the extract having the fibrinolytic activity, three bands are found, protein 1, 2 and 3 having a molecular weight of 30,000, 45,000 and 60,000 Dalton.

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Epithelial-mesenchymal Transition is Associated with Acquired Resistance to 5-Fluorocuracil in HT-29 Colon Cancer Cells

  • Kim, A-Young;Kwak, Jae-Hwan;Je, Nam Kyung;Lee, Yun-hee;Jung, Young-Suk
    • Toxicological Research
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    • v.31 no.2
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    • pp.151-156
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    • 2015
  • 5-Fluorouracil (5-FU) is commonly used for the therapy of colon cancer; however, acquired resistance to 5-FU is a critical barrier to successful treatment and the primary cause of chemotherapy failure. Epithelial-mesenchymal transition (EMT) is a process whereby cells undergo alterations in morphology and molecular characteristics promoting tumor progression and metastasis. Accumulating evidence shows that transition from epithelial to mesenchymal phenotype in cancer cells is associated with their resistance to chemotherapy. However, it is still poorly understood whether EMT is involved in acquired resistance to 5-FU. In this study, we developed an in vitro cell model, 5-FU-resistant HT-29 colon cancer cells, and characterized the differences in cellular morphology and molecular alterations between parental and resistant cells. In accord with mesenchymal-like morphology of 5-FU-resistant HT-29 cells, the expression of the mesenchymal marker fibronectin was significantly increased in these cells in comparision with that in the parental cells. Of interest, we also found a marked increase in the expression of EMT-inducing transcription factors Twist, Zeb1, and Zeb2. Finally, 5-FU-resistant cells showed enhanced migration in comparison with parental HT-29. Taken together, these results indicate that EMT could be associated with 5-FU resistance acquired by HT-29 cells. A specific role of each transcription factor found in this study will require further investigation.

Molecular Prevalence of Cryptosporidium spp. among Companion Birds Kept in Pet Shops in Japan

  • Iijima, Yuko;Itoh, Naoyuki;Phrompraphai, Totsapon;Ito, Yoichi;Kimura, Yuya;Kameshima, Satoshi
    • Parasites, Hosts and Diseases
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    • v.56 no.3
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    • pp.281-285
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    • 2018
  • Cryptosporidium is the most common protozoan that can infect a wide range of animals, including mammals and birds. Avian Cryptosporidium spp. can cause enteric and respiratory diseases which can be fatal in birds and some species are zoonotic. Companion birds have the potential as reservoir due to their close contact with humans. Pet shops are the major source of companion birds. However, few reports are available regarding Cryptosporidium spp. infection among companion birds kept in pet shops. The present study reports the prevalence and molecular characteristics of Cryptosporidium spp. among companion birds kept in pet shops in Japan. A total of 265 fresh fecal samples were obtained from birds kept in 4 pet shops; these birds belonged to 41 species in 3 bird orders. A nested polymerase chain reaction (PCR) assay targeting the small subunit rRNA gene was employed for the detection of Cryptosporidium spp. A total of 24 samples (9.1%) were positive, and Cryptosporidium spp. were detected from all pet shops. The prevalence of Cryptosporidium spp. in each of the bird orders was 6.5% (10/153) in Psittaciformes, 14.4% (13/90) in Passeriformes, and 4.5% (1/22) in Galliformes. Based on sequence analysis, 13 (54.2%) isolates were classified to C. galli, 8 (33.3%) were avian genotype III, and the remaining 3 (12.5%) were C. baileyi. No infection with zoonotic C. meleagridis and no coinfection with multiple Cryptosporidium spp. and/or genotypes were observed. The zoonotic potential of Cryptosporidium spp. infecting companion birds kept in pet shops in Japan is likely to be low.

Characteristics of Urease from Vibrio parahaemolyticus Possessing tah and the Genes Isolated in Korea

  • Kim, Young-Hee;Kim, Jong-Sook
    • Journal of Microbiology
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    • v.39 no.4
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    • pp.279-285
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    • 2001
  • Vibrio parahaemolyticus is a halophilic bacterium associated with seafood gastroenteritis. An unusual strain of Kanagawa-positive urease producing Vibrio parahaemolyticus O1:K1 was isolated from the environment and identified . A polymerase chain reaction assay revealed that this strain harbored both the tdh and the genes. The urease from this strain was studied. Maximum urease production was induced in LB medium containing 0.2% urea, 0.5% glucose, 2% NaCl and pH 5.5 with 6h of culti-vation at 37$\^{C}$ under aeration. Purification of urease was achieved by the process of whole cell lysate, 65% ammonium sulphate precipitation, DEAE-cellulose ion exchange column chromatography, Sepharose CL-6B gel filtration and oxirane activated Sepharose 6B-urea affinity chromatography with 203 fold purification and 2.2% yield. Analysis of the purified enzyme by SDS-PAGE demonstrated the presence of the subunits with a molecular weight of 85kDa, 59kDa, 41kDa and the molecular weight for the native enzyme by nondenaturing PAGE and gel filtration chromatography was 255kDa. The purified urease was stable at pH 7.5 and the opeimal pH in HEPES buffer was 8.0 The enzyme was stable at 60$\^{C}$ for 2 h with a residual activity of 32% . The addition of 10$\mu$M if NiCl$_2$maintained stability for 30 min. The Km value of the purified enzyme was 35.6 mM in urea substrate. The TD$\_$50/(median toxic dose) of the purified urease was 2.5$\mu\textrm{g}$/ml on human leukemia cells.

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Molecular Systematics of Rhizoctonia solani Isolates from Various Crops with AFLP (AFLP를 이용한 Rhizoctonia solani 균주들의 분자계통학적 특성규명)

  • 최혜선;김경수;김희종;이윤수
    • Korean Journal of Microbiology
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    • v.36 no.1
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    • pp.40-45
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    • 2000
  • Rhizoctonia solani Kuhn[Thanatephorus cucumeris (Frank) Donk]is a destrutive soil-borne plant pathogen affecting many agricultural crops worldwide. R. solani is divided into anastomosis groups based on the ability of the hyphae to fuse, and into subgroups based on morphological, physiological characteristics. AG classifications are convenient and useful in identifying primary causal agents of Rhizoctonia diseases, although the mechanism of anastomosis is not fully understood. Beacause of the difficulties, we sought to develop a more direct method for genetic identification and charaterization of R. solani. Twenty nine isolates of R. solani were used for the analysis of genetic relationships among themselves and for rapid anastomosis grouping with AFLP method. All isolates studied were divided into five groups. Isolate 6 was included in AG-3 with 67% genetic similarity. When isolates 3 was compared with 13 and 10 each, they showed more than 84% and 83% similarity, respectively. Isolates 3, 4, 5, 13, and 16 were included in AG-1 with 83% genetic similarity. Isolates 1, 7, and 8 were included in AG-1(IB).

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First record of a marine microalgal species, Chlorella gloriosa (Trebouxiophyceae) isolated from the Dokdo Islands, Korea

  • Kang, Nam Seon;Lee, Jung A;Jang, Hyeong Seok;Kim, Kyeong Mi;Kim, Eun Song;Yoon, Moongeun;Hong, Ji Won
    • Korean Journal of Environmental Biology
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    • v.37 no.4
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    • pp.526-534
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    • 2019
  • Chlorella gloriosa (Chlorellaceae, Trebouxiophyceae) was isolated from seawater off the coast of the Dokdo Islands in Korea. An axenic culture was established using the streak-plate method on f/2 agar media supplemented with antibiotics, allowing identification of the isolate by morphological, molecular, and physiological analyses. The morphological characteristics observed by light and electron microscopy revealed typical morphologies of C. gloriosa species. The molecular phylogenetic inference drawn from the small-subunit 18S rRNA sequence verified that the microalgal strain belongs to C. gloriosa. Additionally, gas chromatography-mass spectrometry analysis showed that the isolate was rich in nutritionally important omega-3 and -6 polyunsaturated fatty acids and high-performance liquid chromatography analysis revealed that the high-value antioxidants lutein and violaxanthin were biosynthesized as accessory pigments by this microalga, with arabinose, galactose, and glucose as the major monosaccharides. Therefore, in this study, a Korean marine C. gloriosa species was discovered, characterized, and described, and subsequently added to the national culture collection.

Studies of Adsorption on the Anionic Surface of $SiO_2$ by Cationic Modified Starches (전분의 양성화 개질을 통한 음이온성 $SiO_2$ 표면에서의 흡착 특성 연구)

  • Han, DongSung;Kim, YuMi;Kim, HanYoung;Chi, GyeongYup;Cho, InShik;Kim, JongDuk
    • Applied Chemistry for Engineering
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    • v.24 no.1
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    • pp.24-30
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    • 2013
  • The adsorption characteristics of cationic starches and starch-oligomers were investigated using the quartz crystal microbalance with dissipation monitoring (QCM-D). The adsorbed amount of modified starches was higher than that of cationic surfactants such as $C_{12{\sim}16}$ trimethylammonium bromide. Cationic starches did not show the tendency depending on the degree of cationic substitution and molecular weight. On the other hand, the softness of the adsorption layer increased with the molecular weight of cationic starches in a viscoelasticity terms. During the adsorption/desorption steps, the amount of adsorbed cationic surfactants was 4~9 times. On the other hand, the difference in the amount of adsorption of all the $C_1$ grafted cationic starches was just 0~50%. In addition, the rigidity of the adsorption layer of cationic surfactant in the desorption step decreased, while, that of cationic starches increased at the same condition.