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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)

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