Peptide selection and antibody generation for the prospective immunorecognition of Cry1Ab16 protein of transgenic maize

- In silico analysis of Cry1A(b)16 toxin of transgenic maize.
- Identification of peptide markers for the biorecognition of transgenic crops.
- T- and B-cell prediction tools to assess linear epitopes of Cry-1-type proteins.
- Conformational epitope prediction using three Cry1A(b)16 computational models.
- Two peptides were selected, synthesised and used for polyclonal antibody production.

The introduction of genes isolated from different Bacillus thuringiensis strains to express Cry-type toxins in transgenic crops is a common strategy to confer insect resistance traits. This work intended to extensively in silico analyse Cry1A(b)16 protein for the identification of peptide markers for the biorecognition of transgenic crops. By combining two different strategies based on several bioinformatic tools for linear epitope prediction, a set of seven peptides was successfully selected as potential Cry1A(b)16 immunogens. For the prediction of conformational epitopes, Cry1A(b)16 models were built on the basis of three independent templates of homologue proteins of Cry1A(a) and Cry1A(c) using an integrated approach. PcH_736-746 and PcH_876-886 peptides were selected as the best candidates, being synthesised and used for the production of polyclonal antibodies. To the best of our knowledge, this is the first attempt of selecting and defining linear peptides as immunogenic markers of Cry1A(b)-type toxins in transgenic maize.