<i>Bacillus thuringiensis</i> subsp. Entomocidus HD9 Cry1Ba4 insecticidal crystal protein: In-silico mutation, cloning, expression, mutation and domain I functional study

Authors

  • Mohd Fadzli Ahmad Faculty of Engineering and Life Sciences, Universiti Selangor, Jalan Timur Tambahan, 46000 Bestari Jaya, Selangor
  • Hasdianty Abdullah Faculty of Engineering and Life Sciences, Universiti Selangor, Jalan Timur Tambahan, 46000 Bestari Jaya, Selangor

DOI:

https://doi.org/10.54987/bstr.v3i2.296

Keywords:

Bacillus thuringiensis, Polymerase Chain Reaction, site- directed mutagenesis, Cry 1Ba4 protein, Plutella xylostella, homology modelling

Abstract

The 3D structure of the insecticidal protein Cry1Ba4 produced by B. thuringiensis subsp. Entomocidus HD-9 was determined using homology modelling. From the model built, we have been able to identify the possible sites for structure modification by site-directed mutagenesis. The mutation was introduced at the conserved region of α-helix 7 by substituting the hydrophobic motif that comprises alanine 216, leucine 217 and phenylalanine 218 with arginine. Wild and mutant Cry1Ba4 genes were cloned into pET200/D-TOPO and expressed in the expression host. The result suggests that mutant Cry1Ba4 protein was less toxic to the larvae Plutella xylostella compared to the wild-type. In conclusion, alteration in the structure of Domain I had left an impact on the toxicity of Cry1Ba4 against P. xylostella.

References

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Published

2015-12-15

How to Cite

Ahmad, M. F., & Abdullah, H. (2015). <i>Bacillus thuringiensis</i> subsp. Entomocidus HD9 Cry1Ba4 insecticidal crystal protein: In-silico mutation, cloning, expression, mutation and domain I functional study. Bioremediation Science and Technology Research (e-ISSN 2289-5892), 3(2), 20–25. https://doi.org/10.54987/bstr.v3i2.296

Issue

Vol. 3 No. 2 (2015)

Section

Articles

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