• Title/Summary/Keyword: Drosophila S2 Cells

Search Result 37, Processing Time 0.024 seconds

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
    • /
    • v.10 no.1
    • /
    • pp.115-118
    • /
    • 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.

  • PDF

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
    • /
    • 2003.10a
    • /
    • pp.149-150
    • /
    • 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)

  • PDF

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
    • /
    • 2003.10a
    • /
    • pp.147-148
    • /
    • 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)

  • PDF

Studies on the wild yeasts in Korea (II) (한국산 야생효모에 관한 연구 2)

  • 박명삼;라철호
    • Korean Journal of Microbiology
    • /
    • v.8 no.3
    • /
    • pp.95-102
    • /
    • 1970
  • From the crops Drosophila collected in Mt. Sokni and Mt.Kyeryong, 7 strains were isolated and then 6 species of wild yeast were identified. 1) Of these six species of wild yeasts two were to be of genus Saccharomyces(Ascosporgenous), two Torulopsis and two Trichosporon (both genuses of Asporogenous). 2) It was found that the fermentation of the wild yeasts isolated from Drosophila was much better than that of any others ; in particular, S. florentinus and S. cerevisiae were good in fermenting maltose. 3) After being cultivated in malt extract agar medium at $25^{\circ}C$ for 3 days, the vegetative cells were found to be big but Torulopsis cells small. 4) It was also observed that the species of yeasts used fro food by Drosophila largely depends on genus and species of Drophila. 5) Of the yeasts isolated from the Drosophila, Trichosporon capitatum and Torulopsis dattila, which has not previously been recorded, were identified. 6) It is believed, therfore, that S.florentinus, powerful in fermenting maltose, will be extremely useful in terms of industrial application.

  • PDF

Effects of Recombinant Baculovirus Infection Conditions on Production of Green Fluorescent Protein in Drosophila S2 Cells (초파리 S2 세포 시스템에서 녹색형광단백질 생산을 위한 재조합 배큘로바이러스의 감염조건들의 영향)

  • Cho, Hye Sook;Kim, Yeon Kyu;Kim, Kyoung Ro;Cha, Hyung Joon
    • Korean Chemical Engineering Research
    • /
    • v.44 no.1
    • /
    • pp.40-45
    • /
    • 2006
  • The baculovirus-insect Drosophila melanogaster S2 cell system combines advantages of conventional baculovirus system and non-lytic S2 cell system because baculoviruses can infect non-permissive cells such as mammalian and Drosophila S2 cells but cannot replicate themselves inside the cells. In the present work, we investigated effects of infection conditions on production of green fluorescent protein (GFP) as a target protein using this baculovirus-S2 cell system. Even though higher MOI and longer baculovirus contact time showed better GFP expression yield during the shorter period, overall protein yield could be lower during the longer period due to the relatively higher cell detachment and lysis (lower cell viability). In addition, maintaining high MOI will be not practical for large-scale cell culture. Therefore, instead of maintaining high MOI, we found that high initial cell number and concentrated (10X) baculovirus volume can confer comparable protein expression even under the moderate MOI condition. Also, we found that the post-infection time that is connected to state of cells after infection was an important factor for production yield.

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
    • /
    • v.12 no.4
    • /
    • pp.563-568
    • /
    • 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.

Suspension culture of Stably Transformed Drosophila melanogaster S2 Cells expressing EGFP and EPO

  • Sohn, Bong-Hee;Lee, Jong-Min;Kim, Yong-Soon;Kang, Pil-Don;Lee, Sang-Uk;Chung, In-Sik
    • Proceedings of the Korean Society of Sericultural Science Conference
    • /
    • 2003.04a
    • /
    • pp.40-40
    • /
    • 2003
  • Recombinant plasmids harboring heterologous genes coding enhanced green fluorescent protein (EGFP) and erythropoietin (EPO) were transfected and expressed in Drosophila melanogaster S2 cells. Stably transformed cell populations expressing EGFP or monkey EPO were isolated after 4 weeks of selection with hygromycin B. (omitted)

  • PDF

An Increased Intracellular Calcium Ion Concentration in Response to Dimethyl Sulfoxide Correlates with Enhanced Expression of Recombinant Human Cyclooxygenase 1 in Stably Transfected Drosophila melanogaster S2 Cells (Dimethyl sulfoxide에 의한 세포내 칼슘이온 농도 증가가 안정적으로 형질 전환된 초파리 S2 세포에서 재조합 사람 cyclooxygenase 1의 발현에 미치는 영향)

  • Chang, Kyung Hwa;Park, Jong-Hwa;Kim, Do Hyung;Chung, Ha Young;HwangBo, Jeon;Lee, Hyun Ho;Lee, Hee-Young;Shon, Dong-Hwa;Kim, Wonyong;Chung, In Sik
    • KSBB Journal
    • /
    • v.27 no.5
    • /
    • pp.313-318
    • /
    • 2012
  • Dimethyl sulfoxide (DMSO) increased the intracellular calcium ion concentration in stably transfected Drosophila melanogaster S2 cells expressing recombinant cyclooxygenase 1 (COX-1). DMSO did not increase the Drosophila NOS (dNOS) transcript level in calcium chelator-treated cells. Expression of recombinant COX-1 due to DMSO was diminished in cells treated with calcium chelators or channel blockers. Our results indicate that an increased intracellular calcium ion concentration due to DMSO is associated with up-regulation of the dNOS gene, leading to enhanced expression of COX-1.

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
    • /
    • v.27 no.5
    • /
    • pp.583-589
    • /
    • 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.

Chromosome Studies on Several Wild Sepcies of Drosophilidae (야생 초파리 수종의 염색체에 관한 연구)

  • 강영선;김영진;방규환
    • The Korean Journal of Zoology
    • /
    • v.7 no.2
    • /
    • pp.13-18
    • /
    • 1964
  • The chromosomes of thirteen wild forms of Drosophila obtained from Kwangnung in Kyunggi Province, Korea were investigated with the ganglion cells of both male and female larvae using the aceto-lactic orcein squashed method. The male chromosome patterns of the species observed in the present study are summarized as follows:

  • PDF