Next Article in Journal
The Unexpected Tuners: Are LncRNAs Regulating Host Translation during Infections?
Previous Article in Journal
Chemodenervation of the Larynx
Article Menu
Issue 11 (November) cover image

Export Article

Open AccessArticle
Toxins 2017, 9(11), 358; https://doi.org/10.3390/toxins9110358

The First Cry2Ac-Type Protein Toxic to Helicoverpa armigera: Cloning and Overexpression of Cry2ac7 Gene from SBS-BT1 Strain of Bacillus thuringiensis

1
School of Biological Sciences, University of the Punjab, Quaid-i-Azam Campus, Lahore 54590, Pakistan
2
Department of Biotechnology, Lahore College for Women University, Lahore 54590, Pakistan
*
Author to whom correspondence should be addressed.
Academic Editor: Anne-Brit Kolstø
Received: 26 August 2017 / Revised: 12 October 2017 / Accepted: 27 October 2017 / Published: 3 November 2017
(This article belongs to the Section Bacterial Toxins)
View Full-Text   |   Download PDF [11256 KB, uploaded 6 November 2017]   |  

Abstract

The Cry (crystal) proteins from Bacillus thuringiensis are known to have toxicity against a variety of insects and have been exploited to control insect pests through transgenic plants and biopesticides. B. thuringiensis SBS BT-1 carrying the cry2 genes was isolated from soil samples in Pakistan. The 2-kb full length cry2Ac gene was cloned, sequenced, and submitted to the EMBL DNA database (Accession No. AM292031). For expression analysis, Escherichia coli DH5α was transformed with the fragment sub-cloned in pET22b expression vector using NdeI and HindIII restriction sites, and later confirmed by restriction endonuclease analysis. To assess the toxicity of Cry2Ac7 protein against lepidopteran and dipteran insects, BL21 (codon plus) strain of E. coli was further transformed with the recombinant plasmid. The 65-kDa protein was expressed in the form of inclusion bodies up to 180 OD units per liter of the medium. Inclusions were washed with a buffer containing 1.5% Triton-X 100 and >90% pure Cry2Ac7 was obtained. The inclusion bodies were dissolved in 50 mM K2CO3 (pH 11.5), dialyzed, and freeze-dried. This freeze-dried protein as well as inclusion bodies were used in bioassays against larvae of Helicoverpa armigera and Musca domestica. The freeze-dried protein was toxic to H. armigera larvae with an LC50 value of 131 ng/mL. However, Cry2Ac7 produced in E. coli did not show any mortality to M. domestica larvae. This is the first report of Cry2Ac protein toxic to H. armigera. View Full-Text
Keywords: Bacillus thuringiensis (Bt); cry2Ac7 gene; cloning; protein expression; bioassays; Helicoverpa armigera; Musca domestica Bacillus thuringiensis (Bt); cry2Ac7 gene; cloning; protein expression; bioassays; Helicoverpa armigera; Musca domestica
Figures

Figure 1

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).

Supplementary material

Share & Cite This Article

MDPI and ACS Style

Saleem, F.; Shakoori, A.R. The First Cry2Ac-Type Protein Toxic to Helicoverpa armigera: Cloning and Overexpression of Cry2ac7 Gene from SBS-BT1 Strain of Bacillus thuringiensis. Toxins 2017, 9, 358.

Show more citation formats Show less citations formats

Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Toxins EISSN 2072-6651 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top