Victor Arokia Doss, D (2022) Cloning and expression of the vegetative insecticidal protein (vip3V) gene of Bacillus thuringiensis in Escherichia coli. Protein Expression and Purification, 26. pp. 82-88.

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Abstract

A genomic libraryof Bacillus thuringiensis var. kurstaki (B.t.k.) was constructed and a positive clone harboring the full-length
gene encoding a novel vegetative insecticidal protein (Vip3V) was characterized. The vip3V gene was subcloned into pET-22b(+)
vector and overexpressed in Escherichia coli to an extent of about 30% of the total protein. While transcription was influenced bythe
T7 promoter of the vector, synthesis of Vip3V in E. coli host occurred from the B.t.k. ribosomal binding site (rbs) found 917 bp
downstream of the insert and not from the E. coli rbs of the vector. The expressed Vip3V protein was found in the soluble and
periplasmic fractions as well as in the inclusion bodies. A simplified anion-exchange chromatographic method for the purification of
Vip3V using step gradient or one-step elution was developed. The purified protein showed broad-spectrum activityagainst some of
the lepidopteran larvae tested and did not show anyactivityagainst the larvae of silkworm (Bombyx mori) and mosquito (Culex
quinquefasciatus). � 2002 Elsevier Science (USA). All rights reserved

Item Type: Article
Uncontrolled Keywords: : Bacillus thuringiensis; Vegetative insecticidal protein; Biopesticide; Biocontrol agent; Overexpression; Secretoryprotein
Depositing User: Mr Team Mosys
Date Deposited: 05 May 2023 09:39
Last Modified: 05 May 2023 09:39
URI: http://ir.psgcas.ac.in/id/eprint/1895

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