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Toxins 2018, 10(5), 193; https://doi.org/10.3390/toxins10050193

A Genomic and Proteomic Approach to Identify and Quantify the Expressed Bacillus thuringiensis Proteins in the Supernatant and Parasporal Crystal

1
ERI de Biotecnología y Biomedicina (BIOTECMED), Department of Genetics, Universitat de València, 46100 Burjassot, Spain
2
Departamento de Biologia Geral, Universidade Estadual de Londrina, Londrina 86057-970, Paraná, Brazil
*
Author to whom correspondence should be addressed.
Received: 13 April 2018 / Revised: 30 April 2018 / Accepted: 7 May 2018 / Published: 10 May 2018
(This article belongs to the Special Issue Insecticidal Toxins from Bacillus thuringiensis)
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Abstract

The combined analysis of genomic and proteomic data allowed us to determine which cry and vip genes are present in a Bacillus thuringiensis (Bt) isolate and which ones are being expressed. Nine Bt isolates were selected from Spanish collections of Bt based on their vip1 and vip2 gene content. As a first step, nine isolates were analyzed by PCR to select those Bt isolates that contained genes with the lowest similarity to already described vip1 and vip2 genes (isolates E-SE10.2 and O-V84.2). Two selected isolates were subjected to a combined genomic and proteomic analysis. The results showed that the Bt isolate E-SE10.2 codifies for two new vegetative proteins, Vip2Ac-like_1 and Sip1Aa-like_1, that do not show expression differences at 24 h vs. 48 h and are expressed in a low amount. The Bt isolate O-V84.2 codifies for three new vegetative proteins, Vip4Aa-like_1, Vip4Aa-like_2, and Vip2Ac-like_2, that are marginally expressed. The Vip4Aa-like_1 protein was two-fold more abundant at 24 h vs. 48 h, while the Vip4Aa-like_2 was detected only at 24 h. For Vip2Ac-like_2, no differences in expression were found at 24 h vs. 48 h. Moreover, the parasporal crystal of the E-SE10.2 isolate contains a single type of crystal protein, Cry23Aa-like, while the parasporal crystal from O-V84.2 contains three kinds of crystal proteins: 7.0–9.8% weight of Cry45Aa-like proteins, 35–37% weight of Cry32-like proteins and 2.8–4.3% weight of Cry73-like protein. View Full-Text
Keywords: insect pest control; crop protection; vip proteins; cry proteins insect pest control; crop protection; vip proteins; cry proteins
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Gomis-Cebolla, J.; Scaramal Ricietto, A.P.; Ferré, J. A Genomic and Proteomic Approach to Identify and Quantify the Expressed Bacillus thuringiensis Proteins in the Supernatant and Parasporal Crystal. Toxins 2018, 10, 193.

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