Journal of Sericultural and Entomological Science (한국잠사곤충학회지)

Korean Society of Sericultural Science (한국잠사학회)
권호 표지

Construction of the Novel Baculovirus Transfer Vector Using the p10 Gene of BmNPV

BmNPV의 p10 유전자를 이용한 새로운 전이벡터 개발

  • 강석우 (농촌진흥청 잠사곤충연구소) ;
  • 진병래 (농촌진흥청 잠사곤충연구소, 농촌진흥청 잠사곤충연구소, 농촌진흥청 잠사곤충연구소, 농촌진흥청 잠사곤충연구소, 서울대학교 농업생명과학대학)
  • Published : 1997.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

To develope the novel baculovirus transfer vector, the p10 gene was cloned from the Bombyx mori nuclear polygedrosis virus (BmNPV) vB2 strain isolated from the B. mori larvae of sericultural farms. The novel transfer vector was constructed by using the p10 gene of BmNPV vB2 strain was 210 bp. The TAAG sequence at the -71 bp of upstream from translation initiator ATG and two polyadenylation signal site at the downstream from terminator TAA were also detected in the p10 gene. The 5' and 3' flanking region of the p10 gene amplified by PCR was cloned into pBluescriptII SK(+) and then transfer vector pBm10 was construceted. The 7.9 kb pBm10 was analysed by restriction enzymes and the map was confirmed. In order to determine the expression of foreign gene of pBm10, $\beta$-galactosidase gene was inserted in the SmaI site of foreign gene cloning site of pBm10. The pBm10 containing $\beta$-galactosidase gene was cotranfected wth genomic DNA of BmNPV vB2 into BmN-4 cells. The recombinant baculovirus expressing $\beta$-galactosidase was also produced polygedra in the infected cells. The results indicated that pBm10 is functional, suggesting that in the baculovirus expression vector system, the recombinant virus produced by pBm10 was effective by oral infection for the producing recombinant proteins in in vivo expression.

Keywords

References

  1. Annu. Rev. Entomol. v.35 Baculovirus diversity and molecular biology Blissard, G. W.;Rohrmann, G. F.
  2. J. Virol. v.47 Acquisition of host cell DNA sequences by baculoviruses: relationship between host DNA insertions and FP mutants of Autographa californica and Galleria mellonella nuclear polyhedrosis viruses Freser, M. J.;Smith, G. E.;Summers, M. D.
  3. J. Gen. Virol. v.75 The p10 gene of natural isolates of Bombyx mori nuclear polyhedrosis virus encodes a trunked protein with an Mr of 7700 Hu, N. T.;Lu, Y. F.;Hashimoto, Y.;Maeda, S.;Hou, R. F.
  4. J. Seric. Sci. Jpn. v.53 A bombyx moricell line susceptible to a nuclear polyhedrosis virus Inoue, H.;Mitsuhashi, J.
  5. Biotechniques v.14 A method for producting recombinant baculovirus expression vectors at high frequency Kitts, P. A.;Possee, R. D.
  6. Virology v.139 Nucleotide sequence of the p10 polypeptide gene of Autographa californica nuclear polyhedrosis virus Kuzio, J.;Rohel, D. Z.;Curry, C. J.;Krebs, A.;Carstens, E. H.;Faulkner, P.
  7. Invertebrate cellsystem and applications Maeda, S.
  8. Proc. Natl. Acad. Sci. U.S.A. v.92 Messenger RNA polyadenylation:A universal modofication Manley, J. L.
  9. Baculovirus expression vector-A laboratory manual O'Reilly, D. R.;Miller, L. K.;Luckow, V. A.
  10. Mol. Cell. Biol. v.4 Strong and regulated expression of Escherichia coli β-galactosidase in insect cells with a baculovirus vector Pennock, M. D.;Shoemaker, C.;Miller, L. K.
  11. A lacoratory manual (2nd ed.) Molecular cloning Sambrook, J.;Fritsch, E. F.;Maniatis, T.
  12. Proc. Natl. Acad. Sci. U.S.A. v.74 Nucleotide sequencing with chain-terminating inhibitors Sanger, F.;Nicklen, S.;Coulson, A. R.
  13. J. Gen. Virol. v.68 Immunogold detection of polyhedrin, p10 and virion antigens in Autographa californica nuclear polyhedrosis virus-infected S. frugiperdacells Van der Wilk, F.;Van Lent, J. W. M.;Vlak, J. M.
  14. J. Gen. Virol. v.74 Functional domains of the p10 protein of Autographa californica nuclear polyhedrosis virus Van Oers, M. M.;Flipsen, J. T. M.;Reusken, C. B. E. M.;Sliwinsky, E. L.;Goldbach, R. W.;Vlak, J. M.
  15. J. Gen. Vriol. v.70 A cylifornica nuclear polyhedrosis virus p10 gene function using insertion/deletion mutants Williams, G. V.;Rohel, D. Z.;Kuzio, J.;Faulkner, P.
  16. J. Gen. Vriol. v.74 Nucleotide sequence and transcriptional analysis of the p10 gene of Spodoptera exigua nuclear polyhedrosis virus Zuidema, D.;Van Oers, M. M.;Van Strien, E. A.;Caballero, P. C.;Klok, E. J. ;Goldvach, R. W.;Vlak, J. M.