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

Mosibudi Thabiki Malomane, Kulsum Kondiah, Mahloro Hope Serepa-Dlamini

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

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.

Original languageEnglish
Article number000426
JournalAccess Microbiology
Volume5
Issue number1
DOIs
Publication statusPublished - 2023

Keywords

  • Escherichia coli BL21(DE3)
  • insecticidal toxin
  • Pantoea ananatis strain MHSD5
  • pET SUMO
  • tccZ gene
  • toxin complex

ASJC Scopus subject areas

  • Microbiology
  • Microbiology (medical)

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