Molecular and General Genetics MGG

, Volume 208, Issue 3, pp 384–389 | Cite as

Cloning and expression of 130-kd mosquito-larvicidal δ-endotoxin gene of Bacillus thuringiensis var. Israelensis in Escherichia coli

  • Chanan Angsuthanasombat
  • Wipa Chungjatupornchai
  • Sunee Kertbundit
  • Plernpis Luxananil
  • Chatri Settasatian
  • Prapon Wilairat
  • Sakol Panyim
Article
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Summary

Five recombinant E. coli clones exhibiting toxicity to Aedes aegypti larvae were obtained from a library of 800 clones containing XbaI DNA fragments of 110 kb plasmid from B. thuringiensis var. israelensis. All the five clones (pMU 14/258/303/388/679) had the same 3.8-kb insert and encoded a major protein of 130 kDa which was highly toxic to A. aegypti larvae. Three clones (pMU 258/303/388) transcribed the 130 kD a gene in the same direction as that of lac Z promoter of pUC12 vector whereas the transcription of the other two (pMU 14/679) was in the opposite direction. A 1.9-kb fragment of the 3.8 kb insert coded for a protein of 65 kDa. Partial DNA sequence of the 3.8 kb insert, corresponding to the 5′-terminal of the 130 kDa gene, revealed a continuous reading frame, a Shine-Dalgarno sequence and a tentative 5′-regulatory region. These results demonstrated that the 3.8 kb insert is a minimal DNA fragment containing a regulatory region plus the coding sequence of the 130 kDa protein that is highly toxic to mosquito larvae.

Key words

B. thuringiensis δ-endotoxin gene 130-kDa mosquito-larvicidal protein 
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Copyright information

© Springer-Verlag 1987

Authors and Affiliations

  • Chanan Angsuthanasombat
    • 1
  • Wipa Chungjatupornchai
    • 1
  • Sunee Kertbundit
    • 1
  • Plernpis Luxananil
    • 1
  • Chatri Settasatian
    • 1
  • Prapon Wilairat
    • 1
  • Sakol Panyim
    • 1
  1. 1.Center for Molecular Genetics-Genetic Engineering and Department of Biochemistry, Faculty of ScienceMahidol UniversityBangkokThailand

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