Journal of Sericultural and Entomological Science (한국잠사곤충학회지)

Korean Society of Sericultural Science (한국잠사학회)
권호 표지

Expression of the FLP recombinase of the 2 $\mu$m plasmid of yeast in the cultured cells of Bombyx mori using a transient expression vector

Yeast 2 $\mu$m 플라스미드 유래 FLP recombinase 유전자의 곤충 배양세포내 발현

  • 강석우 (농촌진흥청 잠사곤충연구소) ;
  • 윤은영 (농촌진흥청 잠사곤충연구소, 농촌진흥청 잠사곤충연구소, 농촌진흥청 잠사곤충연구소, 경북대학교 농과대학, 서울대학교 농업생명과학대학)
  • Published : 1997.06.01
Download PDF
⟨ Previous Next ⟩

Abstract

In order to express the FLP recombinase in B. mori cultured cell line, BmN-4, transient expression system using a heat shock protein gene (hsp70) promoter of Dorosophilla melnogaster was constructed. This vector was designated as pHsSV. Activity strength of the hsp70 promoter was compared with that of immediate early gene (IE-1) and polyhedrin gene of BmNPV employing the E. coli $\beta$-galactosidase gene as a reporter gene. The result showed that the pHs $\beta$-gal plasmid vector expressed the $\beta$-galactosidase at 2nd and 3rd day after the transfer of plasmid DNA into BmN-4 cells, which was similar to that of pIE1 $\beta$-gal vector, but different from that of a recombinant virus, vBm $\beta$-gal. For the construction of FLP recombinase transient expression vector, the FLP recombinase gene was cloned by polymerase chain reaction technique. To express the FLP recombinase, this gene was inserted into pHsSV plasmid vector, under the control of the hsp70 promotor, and tranfected in BmN-4 cells. The expressed FLP recombinase was estimated at 44kDa on a 12.5% SDS-PAGE.

Keywords

References

  1. EMBO. J. v.5 The integrase family of sitespecific recombinases: regional similarities and global diversity Argos, P. A.;Landy, K.;Abremski, J. B.;Egan, E. H.;Lindquist, R. H.;Hoess, M. L.;Kahn, S. V. L.;Manayana, L. S.;Pierson Ⅲ;Sternberg, N.;Leong, J. M.
  2. J. Biol. Chem. v.260 The FLP protein of the 2-micron plasmid of yeast. Purification of the protein from Escherichia coli cells expressing the cloned FLP gene Babineau, D.;Vetter, D.;Andrews, B. J.;Gronostajski, R. M.;Proteau, G. A.;Beatty, L. G.;Sadowski, P. D.
  3. Proc. Natl. Acad. Sci. U.S.A. v.78 Integration, transcription and control of a Drosophila heat shock gene in mouse cells Corces, V.;Pellicer, A.;Axel, R.;Meselson, M.
  4. Appl. Environ. Microbiol. v.58 Insecticidal properties of genetically engineered baculoviruses experties of genetically engineered baculoviruses expressing an insect juvenile hormone esterase gene Eldridge, R.;O'Reilly, D. R.;Hammock, B. D.;Miller, L. K.
  5. J. Mol. Biol. v.225 Mutagenesis of a conserved region of the gene encoding the FLP recombinase of Saccharomyces cerevisiae. A role for arginine 191 in binding and ligation Friesen, H.;Sadowski, P. D.
  6. Cell v.59 The FLP recombinase of yeast catalyzes site-specific recombination in the Drosophila genome Golic, K. G.;Lindquist, S.
  7. Nature v.286 Nucleotide sequence of the yeast plasmid Hartley, J. L.;Donelson, J. E.
  8. J. Gen. Virol. v.71 Identifying the RNA polymerases that synthesize specific transcripts of the autographa californica nuclear polyhedrosis virus Huh, N. E.;Weaver, R. F.
  9. Cell v.21 Sequence of three copies of the gene for the major Dorosophila heat shock induced protein and their flanking regions Ingolia, T. D.;Craig, E. A.;McCarthy, B. J.
  10. Proc. Natl. Acad. Sci. U.S.A. v.82 Two-micrometer circle site-specific recomnination: The minimal substrate and the possible role of flanking sequences Jayaam, M.
  11. Trends Genet. v.9 Site-specific recombinases: tools for genome engineering Kilby, N. J.;snaith, M. R.;Murray, J. A. H.
  12. Cytotechnology v.8 High level expression of a frog α-amidating enzyme, AE-Ⅱ, In cultured cells and silkworm using a BOMbyx mori nuclear polyhedrosis virus expression vector Kobayashi, J.;Imanishi, S.;Inoue, H.;Ohsuye, K.;Yamaichi, K.;Tsuruoka, N.;Tanaka, S.
  13. Nature v.227 Cleavage of structual proteins during the assembly of bacteriophage T4 Laemmlin U. K.
  14. Invertebrate cell system and applications Maeda, S.
  15. Biochem. Biophys. Res. Commun. v.165 Increased insecticidal effect by a recombinant baculovirus carrying a synthetic diuretic hormone gene Maeda, S.
  16. Virology v.184 Insecticidal effects of an insect-specific neurotoxin expressed by a recombinant baculovirus Maeda, S.;Volrath, S. L.;Hanzlik, T. N.;Hrper, S. A.;Majima, K.;Maddox, D. W.;Hammock, B. D.;Fowler, E.
  17. FEBS Lett. v.197 Stable transformation of Drosophila Kc cells to antibiotic resistance with the bacterial neomycin resistance gene Maisobhaute, C.;Echalier, G.
  18. Proc. Natl. Acad. Sci. U.S.A. v.92 Messenger RNA polyadenylation: A universal modification Manley, J. L.
  19. Bio.Technology v.9 Development of a recombinant baculovirus expressing an insect-selective neurotoxin: potential for pest control McCutchen, B. F.;Choudary, P. V.;Crenshaw, R.;Maddox, D.;Kamita, S.G.;palekar, N.;Volrath, S.;Fowler, E.;Hammock, B. D.;Maeda, S.
  20. Bio/Technology v.13 Transovarian transmission of a foreign gene in the silkworm, Bombyx mori, by Autographa californica nuclear polyhedrosis virus Mori, H.;Yamao, H.;Nakazawa, H.;Sugahara, Y.;Shirai, N.;Matsubara, F.;Sumida, H.;Imamura, T.
  21. Science v.251 Recombinase mediated gene activation and sitespecific integration in mammalian cells O'Gorman, S.;Fox, D. T.;Wahl, G. M.
  22. Dev. Biol. v.28A Basic conditions for the drug selection and transient gene expression in the cultured cell line of Bombyx mori, In vitro cell Okano, K.;Miyajima, N.;Takada, N.;Kobayashi, M.;Maekawa, H.
  23. Baculovirus expression vectors-A laboratory manual O'Reilly, D R.;Miler, L. K.;Luckow, V. A.
  24. Cell v.37 A Drosophila RNA polymerase Ⅱ transcription factor binds to the regulatory site of an hsp70 gene Parker, C. S.;Topol, J.
  25. Mol. Cell. Biol. v.8 Step-arrest mutants of FLP recominase: implications for the catalytic mechanism of DNA recombination Parsons, R. L.;Prsad, R. V.;Harshey, R. M.;Jayaram, M.
  26. Cell v.30 A regulatory upstream promoter element in the Drosophila hsp70 heat shock gene Pelham, H. R. B.
  27. Mol. cell. Biol. v.5 Drosophila melanogaster cells with a diminant selectable marker Rio, D. C.;Rubinm G. M.
  28. Molecular Cloning, A laboratory manual(2nd ed.) Sambrook, J.;Fritsch, E. F.;maniatis, T.
  29. Proc. Natl. Acad. Sci. U.S.A. v.82 The FLP recombinase of the yeast 2-μm plasmid: Characterization of its recombination site Senecoff, J. E.;Bruckner, R. C.;Cox, M. M.
  30. Nature v.352 Construction of an improved baculovirus insecticide containing an insect specific toxin gene Stewart, L. M. D.;Hirst, M.;Ferber, M. L.;Merryweather, A. T.;Cayley, P. J.;Possee, R. D.
  31. Bombyx mori, Jap. J. Genetics v.65 Transient expression of CAT genes injected into early embryos of the domesticated silkworm Tamura, T.;Kanda, T.
  32. Nature v.352 Insect paralysis baculovirus-mediated expression of a miteneurotoxin gene Tomalski, M. D.;Miller, L. K.
  33. Bio/Technology v.10 Expression of a paralytic neurotoxin gene to improve inscet baculoviruses as biopesticides Tomalski, M. D.;Miller, L. K.
  34. J. Gen. Virol. v.71 Expression of cauliflower mosaic virus gene Ⅰ in insect cells using a novel polyhedrin-based baculovirus expression vector Zuidema, D.;Schouten, A.;Usmany, M.;Maule, A. J. Belsham, G. J.;Roosien, J.;Klinge-Roode, E. C.;Van Lent, J. W. M.;Vlak, J. M.