• Title/Summary/Keyword: melanogaster

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Analysis of Two Promoters that Control the Expression of the GTP cyclohydrolase I Gene in Drosophila melanogaster

  • Byun, Jaegoo;Yoon, Jaeseung;Baek, Kwanghee
    • Molecules and Cells
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    • v.27 no.5
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    • pp.583-589
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    • 2009
  • GTP cyclohydrolase I (GTPCH) is a key enzyme in the de novo synthesis of tetrahydrobiopterin. Previously, the Drosophila melanogaster GTPCH gene has been shown to be expressed from two different promoters (P1 and P2). In our study, the 5'-flanking DNA regions required for P1 and P2 promoter activities were characterized using transient expression assay. The DNA regions between -98 and +31, and between -73 and +35 are required for efficient P1 and P2 promoter activities, respectively. The regions between -98 and -56 and between -73 and -41 may contain critical elements required for the expression of GTPCH in Drosophila. By aligning the nucleotide sequences in the P1 and P2 promoter regions of the Drosophila melanogaster and Drosophila virilrs GTPCH genes, several conserved elements including palindromic sequences in the regions critical for P1 and P2 promoter activities were identified. Western blot analysis of transgenic flies transformed using P1 or P2 promoter-lacZ fusion plasmids further revealed that P1 promoter expression is restricted to the late pupae and adult developmental stages but that the P2 promoter driven expression of GTPCH is constitutive throughout fly development. In addition, X-gal staining of the embryos and imaginal discs of transgenic flies suggests that the P2 promoter is active from stage 13 of embryo and is generally active in most regions of the imaginal discs at the larval stages.

Comparative Studies on Genetic Variabilities of Second Chromosomes in Sasang Natural and Experimental Populations of Drosophila melanogaster (자연과 실험집단내 Drosophila melanogaster 제 2 염색체의 유전적 변이성에 대한 비교 연구)

  • Son, Seong-Gon;Choe, Yeong-Hyeon;Lee, Won-Ho
    • Journal of Environmental Science International
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    • v.3 no.4
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    • pp.297-304
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    • 1994
  • The genetic variabilities of second chromosomes concealed Sasang natural and experimental populations of Drosophila melanogaster have been analyzed. The experimental population was composed of D. melanognter which had the lethal-free second chromosome collected from Sasang natural population in 1982. The results were as follow; The mean frequencies of deleterious genes were estimated % be 33.33% in Sasang natural population and 31.72% in experimental population. The allelism rates in lethal genes isolated from the natural and experimental populations were calculated to be about 0.95% and 12.28%, respectively. The allelism rates between lethal genes isolated from the natural population and those of the experimental population were calculated to be about 0.01%. The mean values of elimination by frequencies of deleterious genes and allelism rates were 0.0011 in the natural population and 0.0124 in the experimental population. The frequencies of phenotypic sterility of males in the natural and experimental populations were estimated to be 1.49% and 1.36%, respectively. The frequencies of genotypic sterility of females and males were estimated to be 0.90% and 1.80% in the natural population, and that of males was 2.38% in the experimental population.

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A Study of Amylase Isozymes in Five Strains of Drosophila melanogaster in Korea (한국산 5계통의 초파리 (Drosophila melanogaster)의 Amylase Isozyme에 관한 연구)

  • Chung, Yong-Jai;Park, Kyung-Sook
    • The Korean Journal of Zoology
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    • v.17 no.3
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    • pp.117-126
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    • 1974
  • The amylase isozyme of the five strains of D. melanogaster in Korea(Yusu, Choongju, Jeju, Sinchon III and Sinchon IV) was examined by the polyacrylamide thin layer gel electrophoresis and the results obtained are presented below: 1. The most strains except the Sinchon IV show only one band $(Amy^1)$ in the zymograms. This implies that those strains may be homogeneous for amylase constitutions. 2. The Sinchon IV strain exhibits varisble band patterns (mostly $Amy^1,2$ and $Amy^1,4$) suggesting that this strain may be of heterogeneous amylase constitutions. 3. The $Amy^1$ strain may be the commonest one in the Korean D. melanogaster populations as in other countries. 4. The results of hybridization studies are hard to interpret. 5. Further studies will be extended to many other strains from various localities of Korea.

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The Genetic Structure of Kimpo Natual Population of Drosophila melanogaster (Drosophila melanogaster의 김포 자연집단이 유전적 구조)

  • 이택준;김남우
    • The Korean Journal of Zoology
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    • v.33 no.1
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    • pp.6-11
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    • 1990
  • Analysis of genetic structure in Kimpo natural population of Drosophila was carried out by utilizing the deleterious gene on the second chromosome of Drosophila melanogaster. Male flies tested were continuously collected for eight years; in late September 1974 and 1981-1987. The frequency of deleterious gene (lethal plus semilethal) ranged from 27.02% in 1983 to 41.48% in 1987, and the values estimated from the eight years samples are highly signihcent from each other with a homogenety test (X$^2$=52.0157, d.f.=28, P<0.005). Allelic rates ranged from 1.30% in 1981 to 5.03% in 1974. And the effective population size by using the rate of allelism was estimated average at 3, 300 pairs. Elimination rate by homozygous of lethal gene ranged from 0.0004 in 1984 to 0.0019 in 1974, and that is for smaller than mutation rate(0.005) at second chromosome. We suppose that stable frequency (about 20%) lethal genes of D. melanogaster in Kimpo natual population are maintained by invade of P-type mutator factor (P element) versus eliminated in heterozygous and homozygous condition of lethal gene.

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Analysis of deleterious genes in natural and experimental populations of Drosophila melanogaster (Drosophila melanogaster 자연 및 실험집단내의 유해유전자 분석)

  • 이원호;최우영;권은전;박희정
    • Journal of Life Science
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    • v.13 no.3
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    • pp.252-257
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    • 2003
  • The genetic variabilities of second chromosomes of Drosophila melanogaster concealed in Busan natural and experimental populations have been analyzed by the Cy//Pm method. The experimental population was composed of D. melanogaster which had the lethal-free second chromosome, collected Sasang natural population in 1982. The frequencies of deleterious genes were estimated to be 14.3∼25.4% in Busan natural population and 26.5∼27.2% in experimental population. The allelism rates in lethal genes isolated from the natural and experimental populations were calculated to be about 0.76% and 9.76∼14.17%, respectively. The value of elimination by the frequencies of deleterious genes and allelism rate was 0.0106and the effective population size estimated to be about 430 flies at the 6570 days population.

Isolation and Characterization of the Ribosomal Protein 46 Gene in Drosophila melanogaster

    • Animal cells and systems
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    • v.2 no.1
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    • pp.113-116
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    • 1998
  • A cDNA clone coding for ribosomal protein 46 (rp46) which is a component of 60S ribosomal large subunit has been identified from Drosophila melanogaster. A cDNA clone encoding S. cerevisiae rp46 was used as a probe to screen a Drosophila larvae cDNA library. The DNA sequence analysis revealed that the cDNA coding for Drosophils rp46 contains a complete reading frame of 153 nucleotides coding for 51 amino acids. The deduced amino acid sequence showed 71-75% homology with those of other eukaryotic organisms. Northern blot analysis showed that about 1-kb rp46 transcripts are abundant throughout fly development. Whole mount embryonic mRNA in situ hybridization also showed no preferential distribution of the transcripts to any specific region. The chromosomal in situ hybridization revealed that the identified gene is localized at position 60C on the right arm of the second polytene chromosome with a possibility of single copy.

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Dimethylsulfoxide and Sodium Butyrate Enhance the Production of Recombinant Cyclooxygenase 2 in Stably Transformed Drosophila melanogaster S2 Cells

  • Lee, Jong-Min;Sohn, Bong-Hee;Kim, Yong-Soon;Kang, Pil-Don;Lee, Sang-Uk;Chung, In-Sik
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.10a
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    • pp.149-150
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    • 2003
  • The purpose of this experiment is to optimize the yield of the recombinant Cox2 from the stably transformed Drosophila melanogaster S2 cells, using dimethylsulfoxide and sodium butyrale. Materials and Methods : Materials - Cell line : Drosophila melanogaster Schneider 2 (S2) cells - vector pMT/BiP/V5-His and pCoHygro (Invitrogen) (omitted)

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Functional Expression of Recombinant Tumstatin in Stably Transformed Drosophila melanogaster S2 Cells

  • Sohn, Bong-Hee;Kim, Yong-Soon;Kang, Pil-Don;Lee, Sang-Uk;Lee, Jong-Min;Chung, In-Sik
    • Proceedings of the Korean Society of Sericultural Science Conference
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    • 2003.10a
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    • pp.147-148
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    • 2003
  • The purpose of this experiment is to confirm whether the recombinant tumstatin revealed from the stably transformed Drosophila melanogaster S2 cells has in vitro capacity. Materials and Methods : Materials - Cell line : Drosophila melanogaster Schneider 2 (S2) cells - vector pMT/BiP/V5-His and pCoHygro (Invitrogen) (omitted)

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Production of Recombinant Rotavirus Capsid Protein VP7 from Stably Transformed Drosophila melanogaster S2 Cells

  • Park, Jong-Hwa;Chang, Kyung-Hwa;Lee, Youn-Hyung;Kim, Hae-Yeong;Yang, Jai-Myung;Chung, In-Sik
    • Journal of Microbiology and Biotechnology
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    • v.12 no.4
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    • pp.563-568
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    • 2002
  • Stably transformed Drosophila melanogaster 52 cells producing recombinant VP7 were obtained, and recombinant VP7 expression was confirmed by Western blot analysis. The molecular weight of recombinant VP7 expressed in 52 cells was approximately 35.5 kDa, and 75% of the total VP7 produced was present in the medium. Recombinant VP7 contained N-linked glycosylated oligosaccharides. Aprotinin, leupeptin, and polyvinylpyrrolidone did not have any noticeable effect on recombinant VP7 production; however, DMSO and sodium butyrate increased its production by 120% and 60%, respectively.

Expression of Enhanced Green Fluorescent Protein from Stably Transformed Drosophila melanogaster S2 Cells

  • Lee, Jong-Min;Park, Jong-Hwa;Chung, In-Sik
    • Journal of Microbiology and Biotechnology
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    • v.10 no.1
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    • pp.115-118
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    • 2000
  • Recombinant plasmids harboring a heterologous gene coding for the enhanced green fluorescent protein (EGFP) were transfected and expressed in Drosophila melanogaster S2 cells. A stable transformation of polyclonal cell populations expressing EGFP were isolated after 4 weeks of selection with hygromycin B. The recombinant EFGP expressed in transformed S2 cells consisted of a molecular weight of 27 kDa. EGFP expression was also confirmed by fluorometric measurement. The maximum EGFP concentration was about 9.3 mg/I. The present findings demonstrate not only the successful stable expression of EGFP in Drosophuila was about 9.3 mgI. The present findings demonstrate not only the successful stable expression of EGFP in Drosophila S2 cells, but also the use of EGFP as a reporter to analyze gene expression, with its potential of a Drosophila cell expression system for recombinant protein production being an alternative to a baculovirus-insect cell expression system.

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