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

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

Polish Journal of Microbiology
Volume 67 (2018): Issue 1 (January 2018)
By: Pedro Fragoso,  Alicia Armijo,  Doris Gómez,  Claudio Gómez,  Marco Bugueño,  Gittith Sánchez and  Juan Venegas  
Open Access
|Feb 2022

Figures & Tables

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.
DOI: https://doi.org/10.5604/01.3001.0011.6138 | Journal eISSN: 2544-4646 | Journal ISSN: 1733-1331
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Language: English
Page range: 19 - 26
Submitted on: May 16, 2017
Accepted on: Nov 7, 2017
Published on: Feb 21, 2022
Published by: Polish Society of Microbiologists
In partnership with: Paradigm Publishing Services
Publication frequency: 4 issues per year
Keywords:
– Colombian strains,
genes,
larvae,
PCR methods,
biological test
Related subjects:
Life sciences,
Microbiology and virology

© 2022 Pedro Fragoso, Alicia Armijo, Doris Gómez, Claudio Gómez, Marco Bugueño, Gittith Sánchez, Juan Venegas, published by Polish Society of Microbiologists
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

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