본문 바로가기
HOME> 논문 > 논문 검색상세

논문 상세정보

The Homologous Region 3 from Bombyx mori Nucleopolyhedrovirus Enhancing the Transcriptional Activity of Drosophila hsp70 Promoter

Tang, Shun-Ming   (Key Laboratory of Silkworm Biotechnology, of the Ministry of Agriculture, Sericultural Research Institute  ); Yi, Yong-Zhu   (Key Laboratory of Silkworm Biotechnology, of the Ministry of Agriculture, Sericultural Research Institute  ); Zhou, Ya-Jing   (Key Laboratory of Silkworm Biotechnology, of the Ministry of Agriculture, Sericultural Research Institute  ); Zhang, Zhi-Fang   (Key Laboratory of Silkworm Biotechnology, of the Ministry of Agriculture, Sericultural Research Institute  ); Li, Yi-Ren   (Key Laboratory of Silkworm Biotechnology, of the Ministry of Agriculture, Sericultural Research Institute  ); He, Jia-Lu   (Key Laboratory of Silkworm Biotechnology, of the Ministry of Agriculture, Sericultural Research Institute  );
  • 초록

    Drosophila melanogaster heat shock protein 70 gene promoter (Dhsp70p) is widely used in transgenic insect to drive exogenous gene, and the homologous region 3 from Bombyx mori nucleopolyhedrovirus (BmNPVhr3) functions as an enhancer for several promoters. To test whether BmNPVhr3 can enhance the Dhsp70ps transcriptional activity, the reporter plasmids, which contain the Dhsp70p, the reporter $\beta$ -galactosidase gene with SV40 terminator and BmNPVhr3 fragment, are constructed and transfected into the insect cell lines (Bm-N cells and Sf-21 cells) by lipofectin-mediated method. The results from the transient expression assay show that BmNPVhr3 significantly increases transcriptional activity of Dhsp70p both under the normal condition and under the heat-shock treatment, although the effects are significantly different between in Bm-N cells and in sf-21 cells. The enhancing behavior of BmNPVhr3 on the Dhsp70p is in an orientation-independent manner. Meanwhile, the effects of heat-shock treatment on Dhsp70p alone or Dhsp70p/BmNPVhr3 combination present no significant difference, indicating that BmNPVhr3 only enhances the transcriptional activity of Dhsp70p, but cant alter its characteristic of the response to the heat-shock stress. The above results suggest that the Dhsp70p/BmNPVhr3 combination is more effective one to drive exogenous gene for transgene or stable cell expression system in insects.


  • 주제어

    Heat shock protein 70 gene .   Promoter.  

  • 참고문헌 (31)

    1. Hung, J. J., T. J. Cheng, M. D. Chang, K. D. Chen, H. L. Huang and Y. K. Lai (1998) Involvement of heat shock elements and basal transcription elements in the differential induction of the 70-kDa heat shock protein and its cognate by cadmium chloride in 9L rat brain tumor cells. J. Cell Biochem. 71, 21-35 
    2. Idahl, L. A., P. E. Sandatom and J. Sehlin (1986) Measurements of serum glucose using the luciferin/luciferase system and a liquid scintillation spectrometer. Analyt. Biochem. 155, 177-181 
    3. Lo, H. R., C. C. Chou, T. Y. Wu, J. P. Yuen and Y. C. Chao (2002) Novel baculovirus DNA elements strongly stimulate activities of exogenous and endogenous promoters. J. Biol. Chem. 277, 5256-5264 
    4. Lu, M., P. J. Farrell, R. Johnson and K. latrou (1997) A baculovirus (Bombyx mori nuclear polyhedrosis virus) repeat element functions as a powerful constitutive enhancer in transfected insect cells. J. Bioi. Chem. 272, 30724-30728 
    5. Ricchi, A., G. Cardu, B. Lettieri, D. E. Fusar-Poli, C. Tacchini, A. Bernelli-Zazzera and G. G. Corbucci (2001) Biomolecular and biochemical response of myocardial cell to ischemia and reperfusion in the course of heart surgery. J. Cardiovasc. Surg. 42, 605-610 
    6. Shen, X. J., Y. Z. Yi, S. M. Tang, Z. F. Zhang, Y. R. Li, J. L. He and X. F. Wu (2004) Characterization of ecdysteroid UDP-glucosyltransferase gene promoter from Bombyx mori nucleopolyhedrovirus. Int. J. Indust. Entomol. 8, 169-174 
    7. Viswanathan, P., B. Venkaiah, M. S. Kumar, S. Rasheedi, S. Vrati, M. D. Bashyam and S. E. Hasnain (2003) The homologous region sequence (hr1) of Autographa californica multinucleocapsid polyhedrosis virus can enhance transcription from non-baculoviral promoters in mammalian cells. J. BioI. Chem. 278, 52564-52571 
    8. Zhang, Z. F., Y. Zhang, H. S. Lu and X. F. Wu (1995) Structure and function of DNA replication origin hr3 of Bombyx mori nuclear polyhedrosis virus. Sci. China Series B. 25, 949-955 
    9. Zuidema, D., A. Schouten, M. Usmany, A. J. Maule, G. J. Belsham, J. Roosien, E. C. Klinge-Roode, J. W. van Lent and J. M. Vlak (1990) Expression of cauliflower mosaic virus gene I in insect cells using a novel polyhedrin-based baculovirus expression vector. J. Gen. Virol. 71, 2201-2209 
    10. Zhao, Y. G. and P. Eggleston (1999) Comparative analysis of promoters for transient gene expression in cultured mosquito cells. Insect Mol. Biol. 8, 31-38 
    11. Cotto, J. J. and R. I. Morimoto (1999) Stress-induced activation of the heat-shock response: cell and molecular biology of heat-shock factors. Biochem. Soc. Symp. 64,105-118 
    12. Tang, S. M., Y. Z. Yi, X. J. Shen, Z. F. Zhang, Y. R. Li and J. L. He (2003) Functional analysis of the larval serum protein gene promoter from silkworm, Bombyx mori. Chin. Sci. Bull. 48, 2611-2615 
    13. Zhou, Y. J., Y. Z. Yi, Z. F. Zhang, J. L. He, Y. X. Zhang and X. F.Wu (2003) Promoter activities in the baculovirus envelope glycoprotein gp64 gene. Acta Biochim. Biophys. Sin. 35, 18-26 
    14. Summers, M. D. and G. E. Smith (1987) A manual of methods for baculovirus vectors and insect cell culture procedures. Texas Agricultural Experiment Station Bulletin, No. 1555 
    15. Zhou, Y. J. (2002) Effects of insect hormones and CTAB on the baculovirus replication and related gene promoter activity. Ph. D. Thesis, Engineering University of East China 
    16. Kimura, R. H., P. V. Choudary and C. W. Schmid (1999) Silkworm Bm1 SINE RNA increases following cellular insults. Nucleic Acids Res. 27, 3380-3387 
    17. Sambrook, J., E. F. Fritsch and T. Maniatis (1989) Molecular Cloning: A Laboratory Manual. 2nd ed, Cold Spring Harbor Laboratory Press, New York 
    18. Feder, M. E. and G. E. Hofmann (1999) Heat-shock proteins, molecular chaperones, and the stress response: evolutionary and ecological physiology. Annu. Rev. Physiol. 61, 243-282 
    19. Huynh, C. Q. and H. Zieler (1999) Construction of modular and versatile plasmid vectors for the high-level expression of single or multiple genes in insects and insect cell lines. J. Mol. BioI. 288, 13-20 
    20. Moos, M. (1995) Analysis of proteins; in Short Protocols in Molecular Biology. Ausubel, F. M., R. Brent, R. E. Kingston, D. D. Moore, J. G. Seidman, J. A. Smith and K. Struhl (eds.), pp. 10.1.4-10.1.5. John Wiley & Sons, Inc 
    21. Schmidt, M., T. Heimberger, P. Gruensfelder, G. Schler and F. Hoppe (2004) Inducible promoters for gene therapy of head and neck cancer: an in vitro study. Eur. Arch. Otorhinolaryngol. 261, 208-215 
    22. Lacoste, A., M. C. De Cian, A. Cueff and S. A. Poulet (2001) Noradrenaline and alpha-adrenergic signaling induce the hsp70 gene promoter in mollusc immune cells. J. Cell Sci. 114, 3557-3564 
    23. Uhlirova, M., M. Asahina, L. M. Riddiford and M. Jindra (2002) Heat-inducible transgenic expression in the silkmoth Bombyx mori. Dev. Genes Evol. 212, 145-151 
    24. Jeffrey, H. M. (1992) Experiments in Molecular Genetics, 11th printing, Cold Spring Harbor Laboratory Press, New York, pp. 398-404 
    25. Lei, X. D., Y. D. Mi, Z. Y. Yuan, Z. P. Li and X. F. Wu (1994) Expression of luciferase gene in silkworm. Chin. Sci. Bull. 39, 847-849 
    26. Xiao, Q. L., Z. F. Zhang, Y. Z. Yi, J. L. He and X. F. Wu (2001) Functional analysis of helicase gene promoter and homologous region 3 enhancer in Bombyx mori nuclear polyhedrosis virus. Acta Biochim. Biophys. Sin. 33, 525-530 
    27. Beckmann, R. P., L. A. Mizzen and W. J. Weilch (1990) Interaction of hsp70 with newly synthesized proteins: implication for protein folding and assembly. Science 248, 850-853 
    28. Statistical Analysis System Users Guide (1990) Version 6.12, SAS. Institute Inc. Cary. North Carolina: Institute Inc. Cary. North Carolina, Vols. 1.2 (on disc) 
    29. Gomi, S., K. Majima and S. Maeda (1999) Sequence analysis of the genome of Bombyx mori nucleopolyhedrovirus. J. Gen. Virol. 80, 1323-1337 
    30. Lindquist, S. and E. A. Craig (1988) The heat-shock proteins. Annu. Rev. Genet. 22, 631-677 
    31. Yanagida, Y., A. Mizuno, T. Motegi, E. Kobatake and M. Aizawa (2000) Electrically stimulated induction of hsp70 gene expression in mouse astroglia and fibroblast cells. J. Biotechnol. 79, 53-61 

 활용도 분석

  • 상세보기

    amChart 영역
  • 원문보기

    amChart 영역

원문보기

무료다운로드
유료다운로드

유료 다운로드의 경우 해당 사이트의 정책에 따라 신규 회원가입, 로그인, 유료 구매 등이 필요할 수 있습니다. 해당 사이트에서 발생하는 귀하의 모든 정보활동은 NDSL의 서비스 정책과 무관합니다.

원문복사신청을 하시면, 일부 해외 인쇄학술지의 경우 외국학술지지원센터(FRIC)에서
무료 원문복사 서비스를 제공합니다.

NDSL에서는 해당 원문을 복사서비스하고 있습니다. 위의 원문복사신청 또는 장바구니 담기를 통하여 원문복사서비스 이용이 가능합니다.

이 논문과 함께 출판된 논문 + 더보기