Random mutagenesis to enhance the toxicity of Bacillus thuringiensisCcry proteins against Earias vittella (F.)

Insecticidal proteins derived from Bacillus thuringiensis (Bt) are widely utilized in a variety of insect control applications, including sprays and transgenic crops. The development of resistance in pests, on the other hand, can lessen the effectiveness of Bt toxins. In this study, we made efforts...

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Main Authors: Zaheer, Hafsa, Khurshid, Huma, Yunus, Fakhar-Un-Nisa, Manzoor, Farkhanda, Jabeen, Ghazala, Kanwal, Zakia
Format: Article
Language:English
Published: Penerbit Universiti Kebangsaan Malaysia 2023
Online Access:http://journalarticle.ukm.my/23240/
http://journalarticle.ukm.my/23240/1/SB%204.pdf
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author Zaheer, Hafsa
Khurshid, Huma
Yunus, Fakhar-Un-Nisa
Manzoor, Farkhanda
Jabeen, Ghazala
Kanwal, Zakia
author_facet Zaheer, Hafsa
Khurshid, Huma
Yunus, Fakhar-Un-Nisa
Manzoor, Farkhanda
Jabeen, Ghazala
Kanwal, Zakia
author_sort Zaheer, Hafsa
building UKM Institutional Repository
collection Online Access
description Insecticidal proteins derived from Bacillus thuringiensis (Bt) are widely utilized in a variety of insect control applications, including sprays and transgenic crops. The development of resistance in pests, on the other hand, can lessen the effectiveness of Bt toxins. In this study, we made efforts to enhance the toxicity of two cry proteins Cry1Ac and Cry2Aa through random mutagenesis against cotton bollworm (Earias vitella), one of the most destructive cotton pests in Pakistan. Random mutagenesis is an important tool for elucidating protein structure-function relationships and for modifying proteins to enhance or change their characteristics. We focused on whole cry proteins for random mutagenesis through error-prone PCR and constructed a recombinant library of cry proteins. Sequence analysis of eight mutants showed the mutations of 34 different nucleotides in Cry1Ac and Cry2Aa genes. All mutants were spared for toxicity bioassays against 2nd instar larvae of spotted bollworm. Cry1Ac mutant RM1AcM4 (D242E) and Cry2Aa mutants RM2AaM2 (T354A, T492R, F511L, G585E, D606Y) showed enhanced toxicity as compared to proteins without mutation. These two mutants comprise the mutations in domain-II of cry proteins important in specificity determining regions on midgut receptors in insect pests.
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spelling oai:generic.eprints.org:232402024-03-21T04:55:39Z http://journalarticle.ukm.my/23240/ Random mutagenesis to enhance the toxicity of Bacillus thuringiensisCcry proteins against Earias vittella (F.) Zaheer, Hafsa Khurshid, Huma Yunus, Fakhar-Un-Nisa Manzoor, Farkhanda Jabeen, Ghazala Kanwal, Zakia Insecticidal proteins derived from Bacillus thuringiensis (Bt) are widely utilized in a variety of insect control applications, including sprays and transgenic crops. The development of resistance in pests, on the other hand, can lessen the effectiveness of Bt toxins. In this study, we made efforts to enhance the toxicity of two cry proteins Cry1Ac and Cry2Aa through random mutagenesis against cotton bollworm (Earias vitella), one of the most destructive cotton pests in Pakistan. Random mutagenesis is an important tool for elucidating protein structure-function relationships and for modifying proteins to enhance or change their characteristics. We focused on whole cry proteins for random mutagenesis through error-prone PCR and constructed a recombinant library of cry proteins. Sequence analysis of eight mutants showed the mutations of 34 different nucleotides in Cry1Ac and Cry2Aa genes. All mutants were spared for toxicity bioassays against 2nd instar larvae of spotted bollworm. Cry1Ac mutant RM1AcM4 (D242E) and Cry2Aa mutants RM2AaM2 (T354A, T492R, F511L, G585E, D606Y) showed enhanced toxicity as compared to proteins without mutation. These two mutants comprise the mutations in domain-II of cry proteins important in specificity determining regions on midgut receptors in insect pests. Penerbit Universiti Kebangsaan Malaysia 2023 Article PeerReviewed application/pdf en http://journalarticle.ukm.my/23240/1/SB%204.pdf Zaheer, Hafsa and Khurshid, Huma and Yunus, Fakhar-Un-Nisa and Manzoor, Farkhanda and Jabeen, Ghazala and Kanwal, Zakia (2023) Random mutagenesis to enhance the toxicity of Bacillus thuringiensisCcry proteins against Earias vittella (F.). Sains Malaysiana, 52 (11). pp. 3061-3073. ISSN 0126-6039 https://www.ukm.my/jsm/english_journals/vol52num11_2023/contentsVol52num11_2023.html
spellingShingle Zaheer, Hafsa
Khurshid, Huma
Yunus, Fakhar-Un-Nisa
Manzoor, Farkhanda
Jabeen, Ghazala
Kanwal, Zakia
Random mutagenesis to enhance the toxicity of Bacillus thuringiensisCcry proteins against Earias vittella (F.)
title Random mutagenesis to enhance the toxicity of Bacillus thuringiensisCcry proteins against Earias vittella (F.)
title_full Random mutagenesis to enhance the toxicity of Bacillus thuringiensisCcry proteins against Earias vittella (F.)
title_fullStr Random mutagenesis to enhance the toxicity of Bacillus thuringiensisCcry proteins against Earias vittella (F.)
title_full_unstemmed Random mutagenesis to enhance the toxicity of Bacillus thuringiensisCcry proteins against Earias vittella (F.)
title_short Random mutagenesis to enhance the toxicity of Bacillus thuringiensisCcry proteins against Earias vittella (F.)
title_sort random mutagenesis to enhance the toxicity of bacillus thuringiensisccry proteins against earias vittella (f.)
url http://journalarticle.ukm.my/23240/
http://journalarticle.ukm.my/23240/
http://journalarticle.ukm.my/23240/1/SB%204.pdf