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Molecular and Cellular Biochemistry

, Volume 351, Issue 1–2, pp 93–98 | Cite as

Recombinant scorpion insect excitatory toxin BmK IT accelerates the growth of insect Spodoptera frugiperda 9 cells

  • Yuejun FuEmail author
  • Renjia Yang
  • Aihua Liang
  • Chenggang Xu
  • Changchen Hu
Article

Abstract

Several scorpion insect toxins are selectively active on the lepidopterous and dipterous insects. The gene encoding insect excitatory neurotoxin (BmK IT) from the scorpion Buthus martensii Karsch was expressed in Escherichia coli BL21(DE3) at a high level of 3 mg/0.5 L using the prokaryotic expression system pTWIN1. Colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), whole-cell patch-clamp technique and immunofluorescence assays were used to evaluate the toxicity of rBmK IT to insect Spodoptera frugiperda 9 (Sf9) cells and to analyze the potential mechanism of this toxicity. rBmK IT accelerated the growth of Sf9 cells in a dose-dependent manner. Voltage-gating sodium channel activity could not be detected in Sf9 cells using a whole-cell patch-clamp technique. However, immunofluorescence analysis clearly showed co-localization of tetrodotoxin (TTX) and rBmK IT on the Sf9 cell membrane, which demonstrated that rBmK IT could bind to and act on the voltage-gated sodium channels on the Sf9 cells by the high affinity action power. The findings presented in this study are essential for further study of this peptide.

Keywords

Buthus martensii Karsch insect toxin (BmK IT) Toxicity Spodoptera frugiperda 9 cell Sodium channel 

Notes

Acknowledgments

We would like to thank Dr. Spyridon Theofilopoulos (Karolinska Institute, Sweden), for critical comments and linguistic styling of the manuscript. This project was supported by grants from “National Natural Science Foundation of China” (No. 30700534 and No. 30901774), “Natural Science Foundation of Shanxi Province” (2008021039), “Medical Science and Technique Foundation of Shanxi Province (200617)” and the “National High Technology Research and Development Program of China (863 Program)”.

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Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  • Yuejun Fu
    • 1
    Email author
  • Renjia Yang
    • 1
  • Aihua Liang
    • 1
  • Chenggang Xu
    • 1
  • Changchen Hu
    • 2
    • 3
  1. 1.Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of EducationInstitute of Biotechnology, Shanxi UniversityTaiyuanPeople’s Republic of China
  2. 2.Department of NeurosurgeryShanxi Provincial People’s HospitalTaiyuanPeople’s Republic of China
  3. 3.Department of NeurosurgeryZhujiang Hospital, Southern Medical UniversityGuangzhouPeople’s Republic of China

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