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Molecular Characterization of the cry Gene profile of Bacillus thuringiensis Isolated from a Caribbean Region of Colombia

Pedro Fragoso
Pedro Fragoso
, 
Alicia Armijo
Alicia Armijo
, 
Doris Gómez
Doris Gómez
, 
Claudio Gómez
Claudio Gómez
, 
Marco Bugueño
Marco Bugueño
, 
Gittith Sánchez
Gittith Sánchez
 and 
Juan Venegas
Juan Venegas
   | Mar 09, 2018
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Article Category: original-paper
Published Online: Mar 09, 2018
Page range: 19 - 26
Received: May 16, 2017
Accepted: Nov 07, 2017
DOI: https://doi.org/10.5604/pjm-2018-6138
Keywords
© 2021 Pedro Fragoso et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Detection of the cry1 gene in Bacillus thuringiensis DNA samples from a Caribbean region of Colombia.Electrophoresis in 2.5% agarose gels, showing PCR amplification products for the cry1 gene using the primers and protocols described by Ben-Dov et al. (1997). Molecular marker: 100 base pair (bp) DNA ladder (lane 1), without sample (lanes 2, 20). Lanes 3–19: B. thuringiensis samples (Bt-UPC-1 to Bt-UPC-17). Lanes 21, 22: different concentrations of Bt-UPC-1. The Bt DNA samples and the volume analyzed are indicated above the figure. Expected size 274–277 bp (See Material and Methods for more details).
Detection of the cry1 gene in Bacillus thuringiensis DNA samples from a Caribbean region of Colombia.Electrophoresis in 2.5% agarose gels, showing PCR amplification products for the cry1 gene using the primers and protocols described by Ben-Dov et al. (1997). Molecular marker: 100 base pair (bp) DNA ladder (lane 1), without sample (lanes 2, 20). Lanes 3–19: B. thuringiensis samples (Bt-UPC-1 to Bt-UPC-17). Lanes 21, 22: different concentrations of Bt-UPC-1. The Bt DNA samples and the volume analyzed are indicated above the figure. Expected size 274–277 bp (See Material and Methods for more details).

Fig. 2.

Aligment of a cry1 protein coding gene segment of two DNA samples from a Caribbean region of Colombia.The corresponding deduced amino acid sequence is given below each nucleotide sequence. The first sequence corresponds to a segment of an open reading frame of the B. thuringiensis var. kurstaki HD-1 cry1A gene, from base pair 2731 to 3080 (Bthu-cry1A, Genbank code D17518.1). The second and third sequences are PCR products of cry1 amplified from Colombian B. thuringiensis Bt-UPC-1 (Bt1-cry1, GenBank code MG271933) and Bt-UPC-3 (Bt3-cry1, GenBank code MG271934) DNA samples, respectively. The box indicates the two single nucleotide polymorphisms (SNP) between sequences, and the corresponding changes of Serine 965 to Phenylalanine in Bt sequences.
Aligment of a cry1 protein coding gene segment of two DNA samples from a Caribbean region of Colombia.The corresponding deduced amino acid sequence is given below each nucleotide sequence. The first sequence corresponds to a segment of an open reading frame of the B. thuringiensis var. kurstaki HD-1 cry1A gene, from base pair 2731 to 3080 (Bthu-cry1A, Genbank code D17518.1). The second and third sequences are PCR products of cry1 amplified from Colombian B. thuringiensis Bt-UPC-1 (Bt1-cry1, GenBank code MG271933) and Bt-UPC-3 (Bt3-cry1, GenBank code MG271934) DNA samples, respectively. The box indicates the two single nucleotide polymorphisms (SNP) between sequences, and the corresponding changes of Serine 965 to Phenylalanine in Bt sequences.

Fig. 3.

Detection of the cry2 gene in Bacillus thuringiensis DNA samples from a Caribbean region of Colombia.Electrophoresis in 2.5% agarose gels, showing PCR amplification products for the cry2 gene using the primers and protocols described by Ben-Dov et al. (1997). Molecular marker: 100 base pair (bp) DNA ladder (lane M), without sample (lane 0). DNA of B. thuringiensis var. israelensi (Bti), 0.34 μg. Colombian B. thuringiensis samples Bt-UPC-18, Bt-UPC-1 to 19 (lanes 18, 1–19, respectively). The volume analyzed for each sample was 1 μl. Expected size about 700 bp (See Material and Methods for more details).
Detection of the cry2 gene in Bacillus thuringiensis DNA samples from a Caribbean region of Colombia.Electrophoresis in 2.5% agarose gels, showing PCR amplification products for the cry2 gene using the primers and protocols described by Ben-Dov et al. (1997). Molecular marker: 100 base pair (bp) DNA ladder (lane M), without sample (lane 0). DNA of B. thuringiensis var. israelensi (Bti), 0.34 μg. Colombian B. thuringiensis samples Bt-UPC-18, Bt-UPC-1 to 19 (lanes 18, 1–19, respectively). The volume analyzed for each sample was 1 μl. Expected size about 700 bp (See Material and Methods for more details).

Fig. 4.

Detection of the cry4B gene in different volumes of Bacillus thuringiensis DNA samples from a Caribbean region of Colombia.The four electrophoreses were performed in 2.5% agarose gels, showing PCR amplification products for the cry4B gene using the primers and protocols described by Santos et al. (2012). Molecular marker: 100 base pair (bp) DNA ladder (lanes 1, 10, 14 and 25), without sample (lanes 2, 11, 15 and 26). Colombian B. thuringiensis samples (lanes 3–9, 12, 13, 16–23 and 27, 28, respectively). The corresponding Bt-UPC DNA samples and the volume analyzed are given above the figure.
Detection of the cry4B gene in different volumes of Bacillus thuringiensis DNA samples from a Caribbean region of Colombia.The four electrophoreses were performed in 2.5% agarose gels, showing PCR amplification products for the cry4B gene using the primers and protocols described by Santos et al. (2012). Molecular marker: 100 base pair (bp) DNA ladder (lanes 1, 10, 14 and 25), without sample (lanes 2, 11, 15 and 26). Colombian B. thuringiensis samples (lanes 3–9, 12, 13, 16–23 and 27, 28, respectively). The corresponding Bt-UPC DNA samples and the volume analyzed are given above the figure.

Fig. 5.

Alignment of the cry4B segments from the Colombian B. thuringiensis Bt – UPC – 22 sample and from the B. thuringiensis serovar israelensis bacterium.These nucleotide sequences correspond to a segment of 140 bp from B. thuringiensis serovar israelensis cry4B gene coding sequence (cry4B, GenBank code: D00247.1) and the 97 bp nucleotide segment sequenced from Colombian B. thuringiensis DNA sample Bt22 (cry4B-Bt22rc). Below each nucleotide sequence is the corresponding deduced amino acid segment.
Alignment of the cry4B segments from the Colombian B. thuringiensis Bt – UPC – 22 sample and from the B. thuringiensis serovar israelensis bacterium.These nucleotide sequences correspond to a segment of 140 bp from B. thuringiensis serovar israelensis cry4B gene coding sequence (cry4B, GenBank code: D00247.1) and the 97 bp nucleotide segment sequenced from Colombian B. thuringiensis DNA sample Bt22 (cry4B-Bt22rc). Below each nucleotide sequence is the corresponding deduced amino acid segment.
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