Genetic Engineering of Escherichia coli BL21 (DE3) with a codon-optimized insecticidal toxin complex gene tccZ .

Autor:	Malomane MT; Department of Biotechnology and Food Technology, Faculty of Science, University of Johannesburg, Doornfontein Campus, PO Box 17011, Doornfontein 2028, Johannesburg, South Africa., Kondiah K; Department of Biotechnology and Food Technology, Faculty of Science, University of Johannesburg, Doornfontein Campus, PO Box 17011, Doornfontein 2028, Johannesburg, South Africa., Serepa-Dlamini MH; Department of Biotechnology and Food Technology, Faculty of Science, University of Johannesburg, Doornfontein Campus, PO Box 17011, Doornfontein 2028, Johannesburg, South Africa.
Jazyk:	angličtina
Zdroj:	Access microbiology [Access Microbiol] 2023 Jan 24; Vol. 5 (1), pp. acmi000426. Date of Electronic Publication: 2023 Jan 24 (Print Publication: 2023).
DOI:	10.1099/acmi.0.000426
Abstrakt:	A toxin complex consists of a high-molecular-weight group of toxins that exhibits insecticidal activity against insect pests. These toxins are a promising alternative to Bacillus thuringiensis ( Bt ) toxins that have been extensively utilized in insect pest control. Herein, a codon-optimized insecticidal gene ( tccZ ) (381 bp) identified in Pantoea ananatis strain MHSD5 (a bacterial endophyte previously isolated from Pellaea calomelanos ) was ligated into the pET SUMO expression vector and expressed in Escherichia coli BL21 (DE3). We report the success of cloning the tccZ gene into the pET SUMO vector and ultimately the transformation into E. coli BL21 (DE3) competent cells. However, despite conducting a time course of expression as well as isopropyl β-d-1-thiogalactopyranoside (IPTG) dosage optimization to determine optimal conditions for expression, TccZ protein expression could not be detected on Stain-Free and Coomassie-stained SDS-PAGE gels. Competing Interests: The authors declare that there are no conflicts of interest. (© 2023 The Authors.)
Databáze:	MEDLINE
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