• Title/Summary/Keyword: melanogaster

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Construction of the Transgenic Drosophila melanogaster Expressing a Human Megsin Gene. (인간 신장질환 유발인자가 발현하는 형질전환 초파리 구축)

  • Goo, Tae-Won;Kwon, Ki-Sang;Kwon, O-Yu
    • Journal of Life Science
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    • v.17 no.5 s.85
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    • pp.719-722
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    • 2007
  • IgA nephropathy(IgAN) is considered to be a multifactorial disease with genetic and environmental factors contributing to its pathogenesis. The genes involved in susceptibility and progression of the disease have not yet been clearly elucidated. Megsin is an important candidate gene, predominantly expressed in glomerular mesangium and upregulated in IgAN. To understand biological function of megsin, in this work we have produced transgenic D. melanogaster fly over-expressing human megsin(actin-gal4>UAS-Megsin fly). Introduced human megsin was confirmed by RT-PCR and Western blotting, respectively. Its phenotype is melanin deficiency-abdomen and the megsin gene is stably transferred to the next generations.

Functional Characterization of Drosophila melanogaster CYP6A8 Fatty Acid Hydroxylase

  • Sang-A Lee;Vitchan Kim;Byoungyun Choi;Hyein Lee;Young-Jin Chun;Kyoung Sang Cho;Donghak Kim
    • Biomolecules & Therapeutics
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    • v.31 no.1
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    • pp.82-88
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    • 2023
  • Genomic analysis indicated that the genome of Drosophila melanogaster contains more than 80 cytochrome P450 genes. To date, the enzymatic activity of these P450s has not been extensively studied. Here, the biochemical properties of CYP6A8 were characterized. CYP6A8 was cloned into the pCW vector, and its recombinant enzyme was expressed in Escherichia coli and purified using Ni2+-nitrilotriacetate affinity chromatography. Its expression level was approximately 130 nmol per liter of culture. Purified CYP6A8 exhibited a low-spin state in the absolute spectra of the ferric forms. Binding titration analysis indicated that lauric acid and capric acid produced type I spectral changes, with Kd values 28 ± 4 and 144 ± 20 µM, respectively. Ultra-performance liquid chromatography-mass spectrometry analysis showed that the oxidation reaction of lauric acid produced (ω-1)-hydroxylated lauric acid as a major product and ω-hydroxy-lauric acid as a minor product. Steady-state kinetic analysis of lauric acid hydroxylation yielded a kcat value of 0.038 ± 0.002 min-1 and a Km value of 10 ± 2 µM. In addition, capric acid hydroxylation of CYP6A8 yielded kinetic parameters with a kcat value of 0.135 ± 0.007 min-1 and a Km value of 21 ± 4 µM. Because of the importance of various lipids as carbon sources, the metabolic analysis of fatty acids using CYP6A8 in this study can provide an understanding of the biochemical roles of P450 enzymes in many insects, including Drosophila melanogaster.

Physiological Genetic Effects of Monomer and Polymer Containing 5-Fluorouracil on Drosophila Melanogaster (5-플루오로우라실을 포함하는 단량체와 중합체의 노랑초파리에 대한 생리유전학적 영향)

  • Lee, Neung-Ju;Kim, Ik-Sung;Choi, Won-Moon;Ha, Chang-Sik;Cho, Won-Jei
    • Applied Chemistry for Engineering
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    • v.2 no.1
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    • pp.56-63
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    • 1991
  • The monomer, 1-(2-carbomethoxyacryloyl)-5-fluorouracil(CMAFU) was synthesized from trans-${\beta}$-carbomethoxyacryl chloride and 2, 4-bis(trimethylsilyloxy)-5-fluoropyrimidine. A copolymer of CMAFU with methyl methacrylate (MMA)[poly(CMAFU-co-MMA)] was also prepared with 2, 2'-azobisisobutyronitrile in cyclohexanone at $60^{\circ}C$. Physiological genetic effects of 5-fluorouracil(5-FU), CMAFU and poly(CMAFU-co-MMA) on Drosophila melanogaster were investigated by the adult feeding method of Lewis and Bacher. It was found that a physiological genetic effect on the Drosophila melanogaster was considerably weaker for CMAFU and its copolymer than 5-FU.

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Changes of Protein Synthesis during Oogenesis of Drosophila melanogaster (노랑초파리의 난자형성과정에서의 단백질 합성의 변화)

  • 박성순;이양림
    • The Korean Journal of Zoology
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    • v.30 no.1
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    • pp.10-28
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    • 1987
  • Changes of protein synthesis in the isolated egg chambers were studied during oogenesis of Drosophila melanogaster. Protein synthesis did not change much in quantity except that the activity was slightly enhanced at the stages 11 and 12, when new structures such as vitelline membranes and yolk granules were made, but considerably changed in quality during oogenesis. Protein synthesis was believed to occur mainly in the follicle cells, and a number of stage-specific proteins were found to differ at various stages. The observations suggest that proteins synthesized in the cells within chambers as well as those from the outside of the chambers might be directly required for the growth and maturation of oocytes in Drosophila.

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Monoclonal Antibody Recognizing Nervous System Specific Protein of Drosophila melanogaster (초파리 신경계특이적인 단일클론항체의 제작과 그 항원의 국재)

  • 윤춘식
    • Journal of Life Science
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    • v.8 no.5
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    • pp.571-575
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    • 1998
  • The nerve system specific protein of Drosophila melanogaster was produced by using heads of flies as the antigen. The monoclonal antibody 6H6 recognized the disabled molecules that a kind of tyrosine kinase substrate by expres-sion cDNA library screening method. At the same time, the antibody also specifically recognized C-terminal region of disabled protein from 7427 to 8761bp by DNA sequencing. In early embryos, the localization of antigen appeared in the central nerve system. In adult flies, the antigen showed specific localization on the axon of optic nerve, cerebral nerve and thoracic nerve, and they also expressed on the muscular nerve. The molecules of disabled are expected to carry an important function in developing central nerve system. In adult flies, it is suggested that the disabled molecules have a role for muscular nerve as well as neural axon.

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Comparative Induction of Sex-linked lethal mutations in Drosophila melanogaster males by Ethyl methanesulfonate (Drosophila meianogaster에서 Ethyl methanesulfonate에 의한 반성 치사 돌연변이 유발성의 비교)

  • Choi, Yung-Hyun;Yoo, Mi-Ae;Lee, Won-Ho
    • Journal of Environmental Science International
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    • v.4 no.5
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    • pp.157-157
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    • 1995
  • In order to analyze the sex-linked lethal mutagenic effects of ethyl methanesulfonate (EMS) in Drosophila melanogaster, the mutagenicities from the attached-X and Basc method have been detected. The indexes of relative lethal mutations at 2.0 and 4.0mM EMS treated group using the attached-X method were about 1.5 and 2.4 times than that of 1.0mM treated group, respectively. EMS had more pronounced effect in the sperm and spermatid stages in the induction of X-linked lethal mutations during the spermatogenesis. And, in the induction of X-linked recessive lethal mutations from the Basc method, the mutation frequency of 4.0mM EMS treated group as compared with 2.0mM was more than three fold. Two assay systems used in this study can support mutually according to experimental purposes, and the area of its application can be considerably wide.

Comparative Induction of Sex-linked lethal mutations in Drosophila melanogaster males by Ethyl methanesulfonate (Drosophila meianogaster에서 Ethyl methanesulfonate에 의한 반성 치사 돌연변이 유발성의 비교)

  • Choi, Yung-Hyun;Yoo, Mi-Ae;Lee, Won-Ho
    • Journal of Environmental Science International
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    • v.4 no.5
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    • pp.543-549
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    • 1995
  • In order to analyze the sex-linked lethal mutagenic effects of ethyl methanesulfonate (EMS) in Drosophila melanogaster, the mutagenicities from the attached-X and Basc method have been detected. The indexes of relative lethal mutations at 2.0 and 4.0mM EMS treated group using the attached-X method were about 1.5 and 2.4 times than that of 1.0mM treated group, respectively. EMS had more pronounced effect in the sperm and spermatid stages in the induction of X-linked lethal mutations during the spermatogenesis. And, in the induction of X-linked recessive lethal mutations from the Basc method, the mutation frequency of 4.0mM EMS treated group as compared with 2.0mM was more than three fold. Two assay systems used in this study can support mutually according to experimental purposes, and the area of its application can be considerably wide.

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Cucurbitacin B Activates Bitter-Sensing Gustatory Receptor Neurons via Gustatory Receptor 33a in Drosophila melanogaster

  • Rimal, Suman;Sang, Jiun;Dhakal, Subash;Lee, Youngseok
    • Molecules and Cells
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    • v.43 no.6
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    • pp.530-538
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    • 2020
  • The Gustatory system enables animals to detect toxic bitter chemicals, which is critical for insects to survive food induced toxicity. Cucurbitacin is widely present in plants such as cucumber and gourds that acts as an anti-herbivore chemical and an insecticide. Cucurbitacin has a harmful effect on insect larvae as well. Although various beneficial effects of cucurbitacin such as alleviating hyperglycemia have also been documented, it is not clear what kinds of molecular sensors are required to detect cucurbitacin in nature. Cucurbitacin B, a major bitter component of bitter melon, was applied to induce action potentials from sensilla of a mouth part of the fly, labellum. Here we identify that only Gr33a is required for activating bitter-sensing gustatory receptor neurons by cucurbitacin B among available 26 Grs, 23 Irs, 11 Trp mutants, and 26 Gr-RNAi lines. We further investigated the difference between control and Gr33a mutant by analyzing binary food choice assay. We also measured toxic effect of Cucurbitacin B over 0.01 mM range. Our findings uncover the molecular sensor of cucurbitacin B in Drosophila melanogaster. We propose that the discarded shell of Cucurbitaceae can be developed to make a new insecticide.

Changes in Protein Patterns During Oogenesis and Development of Drosophila melanogaster (노랑초파리의 난자형성 및 발생과정중의 단백질 양상의 변화)

  • 이양림;문영화;정숙연
    • The Korean Journal of Zoology
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    • v.24 no.1
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    • pp.27-37
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    • 1981
  • Protein patterns at various stages of pre-and post-fertilization in Drosophila melanogaster were analyzed using polyacrylamide gel electrophoresis. Ovaries at various times after hatching, embryos, larvae and pupae were collected and used for protein analysis. The protein patterns were found to vary qualitatively and quantitatively during oogenesis as well as during embryogenesis and metamorphosis. Yolk proteins, which were found to be synthesized in fat body and transferred into the oocytes, appeared in three bands in the gel in the early stages, but these proteins seemded to be converged into a single band in the late stage of oogenesis. The protein patterns in the post fertilization stages seemed to be abruptly changed at two different stages; between embryonic and larval stages and between the second and third instar stages. The pattern in the pupal stages was not very much different from that of the third instar.

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Prevalence of negative frequency-dependent selection, revealed by incomplete selective sweeps in African populations of Drosophila melanogaster

  • Kim, Yuseob
    • BMB Reports
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    • v.51 no.1
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    • pp.1-2
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    • 2018
  • Positive selection on a new beneficial mutation generates a characteristic pattern of DNA sequence polymorphism when it reaches an intermediate allele frequency. On genome sequences of African Drosophila melanogaster, we detected such signatures of selection at 37 candidate loci and identified "sweeping haplotypes (SHs)" that are increasing or have increased rapidly in frequency due to hitchhiking. Based on geographic distribution of SH frequencies, we could infer whether selective sweeps occurred starting from de novo beneficial mutants under simple constant selective pressure. Single SHs were identified at more than half of loci. However, at many other loci, we observed multiple independent SHs, implying soft selective sweeps due to a high beneficial mutation rate or parallel evolution across space. Interestingly, SH frequencies were intermediate across multiple populations at about a quarter of the loci despite relatively low migration rates inferred between African populations. This invokes a certain form of frequency-dependent selection such as heterozygote advantage. At one locus, we observed a complex pattern of multiple independent that was compatible with recurrent frequency-dependent positive selection on new variants. In conclusion, genomic patterns of positive selection are very diverse, with equal contributions of hard and soft sweeps and a surprisingly large proportion of frequency-dependent selection in D. melanogaster populations.