Skip to main content
Log in

Comparison of six promoters for transient expression of luciferase reporter gene in cultured Bombyx mori cells (BmN)

  • Mechanisms of Cell Proliferation and Differentiation
  • Published:
Russian Journal of Developmental Biology Aims and scope Submit manuscript

Abstract

The Luciferase (luc) reporter gene was used to construct recombinant plasmids containing six different promoters, which are the cytoplasmic actin3 promoter (A3), the fibroin heavy chain promoter (Fib-H), the fibroin light chain promoter (Fib-L), a glycoprotein (P25) promoter, Sericin-1 promoter (Ser-1) and the Bombyx mori nuclear polyhedrosis virus immediate (BmNPV) early protein promoter 1(IE-1), respectively. These recombinant plasmids, which are pGL3-A3-luc, pGL3-Fib-H-luc, pGL3-Fib-L-luc, pGL3-P25-luc, pGL3-Ser-1-luc and pGL3-IE-1-luc had been constructed successfully by restriction enzyme digestion and PCR analysis, and then were transfected into BmN cells to measure the activity of the six promoters to drive luc reporter gene transient expression in cells. Transfection and transcriptional experiment indicated that except pGL3-Fib-L-luc, pGL3-P25-luc, pGL3-Ser-1-luc, others three kinds of recombinant plasmids all transfected BmN cells obviously. The promoters of Fib-H, A3 and IE-1 enhanced the transient expression activity of luc reporter gene in the BmN cells and their activity strengthened sequentially.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Angelichio, M.L., Beck, J.A., Johansen, H., and Ivey-Hoyle, M., Comparison of Several Promoters and Polyadenylation Signals for Use in Heterologous Gene Expression in Cultured Drosophila Cells, Nucleic Acids Res., 1991, vol. 19, pp. 5037–5043.

    Article  PubMed  CAS  Google Scholar 

  • Chevillard, M., Couble, P., and Prudhomme, J.C., Complete Nucleotide Sequence of the Gene Encoding the Bombyx mori Silk Protein P25 and Predicted Amino Acid Sequence of the Protein, Nucleic Acids Res., 1986, vol. 14, pp. 6341–6342.

    Article  PubMed  CAS  Google Scholar 

  • Elnitski, L. and Hardison, R., Efficient and Reliable Transfection of Mouse Erythroleukemia Cells Using Cationic Lipids, Blood Cells Mol. Dis., 1999, vol. 25, pp. 299–304.

    Article  PubMed  CAS  Google Scholar 

  • Gambhir, S.S., Barrio, J.R., Herschman, H.R., and Phelps, M.E., Assays for Noninvasive Imaging of Reporter Gene Expression, Nucl. Med. Biol., 1999, vol. 26, pp. 481–490.

    Article  PubMed  CAS  Google Scholar 

  • Guo, T.Q., Wang, J.Y., Guo, X.Y., Wang, S.P., and Lu, C.D., Transient in vivo Gene Delivery to the Silkworm Bombyx mori by EGT-Null Recombinant AcNPV Using EGFP as a Reporter, Arch. Virol., 2005, vol. 150, pp. 93–105.

    Article  PubMed  CAS  Google Scholar 

  • Hua, G., Qian, B., and Wu, X.F., Transient Expression of β-Galactosidase in Bombyx mori Cell (Bm-N) under the Regulation of Fibroin Gene Promoter, Acta Biochim. Biophys. Sin. (Shanghai), 1994, vol. 26, pp. 549–553.

    CAS  Google Scholar 

  • Kuhn, D.E., Martin, M.M., Feldman, D.S., Terry, A.V., Jr., Nuovo, G.J., and Elton, T.S., Experimental Validation of miRNA Targets, Methods, 2008, vol. 44, pp. 47–54.

    Article  PubMed  CAS  Google Scholar 

  • Leclerc, G.M., Boockfor, F.R., Faught, W.J., and Frawley, L.S., Development of a Destabilized Firefly Luciferase Enzyme for Measurement of Gene Expression, Biotechniques, 2000, vol. 29, pp. 590–598.

    PubMed  CAS  Google Scholar 

  • Lee, J.Y., Kim, S., Hwang do, W., Jeong, J.M., Chung, J.K., Lee, M.C., and Lee, D.S., Development of a Dual-Luciferase Reporter System for in vivo Visualization of microRNA Biogenesis and Posttranscriptional Regulation, J. Nucl. Med., 2008, vol. 49, pp. 285–294.

    Article  PubMed  CAS  Google Scholar 

  • Lewis, B.P., Shih, I.H., Jones-Rhoades, M.W., Bartel, D.P., and Burge, C.B., Prediction of Mammalian microRNA Targets, Cell, 2003, vol. 115, pp. 787–798.

    Article  PubMed  CAS  Google Scholar 

  • Liu, Y., Yu, L., Guo, X., Guo, T., Wang, S., and Lu, C., Analysis of Tissue-Specific Region in Sericin 1 Gene Promoter of Bombyx mori, Biochem. Biophys. Res. Commun., 2006, vol. 342, pp. 273–279.

    Article  PubMed  CAS  Google Scholar 

  • Lu, M., Johnson, R.R., and Iatrou, K., Trans-Activation of a Cell Housekeeping Gene Promoter by the IE1 Gene Product of Baculoviruses, Virology, 1996, vol. 218, pp. 103–113.

    Article  PubMed  CAS  Google Scholar 

  • Matsuno, K., Hui, C.C., Takiya, S., Suzuki, T., Ueno, K., and Suzuki, Y., Transcription Signals and Protein Binding Sites for Sericin Gene Transcription in vitro, J. Biol. Chem., 1989, vol. 264, pp. 18707–18713.

    PubMed  CAS  Google Scholar 

  • Naylor, L.H., Reporter Gene Technology: The Future Looks Bright, Biochem. Pharmacol., 1999, vol. 58, pp. 749–757.

    Article  PubMed  CAS  Google Scholar 

  • Nony, P., Prudhomme, J.C., and Couble, P., Regulation of the P25 Gene Transcription in the Silk Gland of Bombyx, Biol. Cell, 1995, vol. 84, pp. 43–52.

    Article  PubMed  CAS  Google Scholar 

  • Pais, H., Nicolas, F.E., Soond, S.M., Swingler, T.E., Clark, I.M., Chantry, A., Moulton, V., and Dalmay, T., Analyzing mRNA Expression Identifies Smad3 as a microRNA-140 Target Regulated Only at Protein Level, RNA, 2010, vol. 16, pp. 489–494.

    Article  PubMed  CAS  Google Scholar 

  • Shen, X.J., Yi, Y.Z., Tang, S.M., Zhang, Z.F., Li, Y.R., and He, J.L., The Ecdysteroid UDP-Glucosyltransferase Gene Promoter from Autographa californica Multicapsid Nucleopolyhedrovirus, Z. Naturforsch. C, 2004, vol. 59, pp. 749–754.

    PubMed  CAS  Google Scholar 

  • Tamura, T., Thibert, C., Royer, C., Kanda, T., Abraham, E., Kamba, M., Komoto, N., Thomas, J.L., Mauchamp, B., Chavancy, G., Shirk, P., Fraser, M., Prudhomme, J.C., and Couble, P., Germline Transformation of the Silkworm Bombyx mori L. Using a piggyBac Transposon-Derived Vector, Nat. Biotechnol., 2000, vol. 18, pp. 81–84.

    Article  PubMed  CAS  Google Scholar 

  • Tang, S., Zhao, Q., Yi, Y., Zhang, Z., and Li, Y., Homologous Region 3 from Bombyx mori Nucleopolyhedrovirus Enhancing the Transcriptional Activity of Heat Shock Cognate 70-4 Promoter from Bombyx mori and Bombyx mandarina in vitro and in vivo, Biosci. Biotechnol. Biochem., 2005, vol. 69, pp. 1014–10177.

    Article  PubMed  CAS  Google Scholar 

  • Zhang, Y., Zhang, X., Xia, H., Xue, Y., Wang, J., Tian, B., Wei, Z., and Lu, C., Quantitative Analysis of Cytoplasmic Actin Gene Promoter and Nuclear Polyhedrosis Virus Immediate-Early Promoter Activities in Various Tissues of Silkworm Bombyx mori Using Recombinant Autographa californica Nuclear Polyhedrosis Virus as Vector, Acta Biochim. Biophys. Sin. (Shanghai), 2008, vol. 40, pp. 533–538.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Xing Jia Shen.

Additional information

The article is published in the original.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Huang, Y., Wang, J.P., Song, F. et al. Comparison of six promoters for transient expression of luciferase reporter gene in cultured Bombyx mori cells (BmN). Russ J Dev Biol 43, 372–376 (2012). https://doi.org/10.1134/S1062360412060021

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S1062360412060021

Keywords

Navigation