Biotechnology Techniques

, Volume 10, Issue 2, pp 89–92 | Cite as

Production and purification of firefly luciferase inEscherichia coli

  • Xuejun Chen
  • Shiren Ren
  • Zhenhua Jin
  • Shenggeng Zhu
Article

Summary

A genetic recombinant stain ofE.coli were induced to express and secrete firefly luciferase. Cells, when broken by freeze/thawing, gave about 2% of the total soluble protein as luciferase. The luciferase was purified with ammonium sulphate fractionation and gel filtration chromatography giving a luciferase product with high specific activity (106 light units/mg protein). SDS-PAGE of this product showed two active bands at 54 and 50 kDa, which corresponded to the luciferases with and without a signal peptide on their N-terminals. The yield was more than one mg purified enzyme per 100 ml of fermentative liquid.

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Copyright information

© Chapman & Hall 1996

Authors and Affiliations

  • Xuejun Chen
    • 1
  • Shiren Ren
    • 1
  • Zhenhua Jin
    • 2
  • Shenggeng Zhu
    • 1
  1. 1.College of Life Sciences, Peking UniversityBeijingP. R. China
  2. 2.Institute of Developmental biology, Academia SinicaBeijingP. R. China

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