Construction, Cloning, and Overexpression of Staphylococcal Enterotoxin B Gene Synthetic (SEBsyn) in pET-28a(+): Pre-development Bacterial-Toxin Therapy for Cancer

DOI

10.2991/absr.k.220406.065How to use a DOI?

Keywords

Bacterial toxin; Cancer; pET-28a(+); SEB; Staphylococcus aureus

Abstract

Staphylococcal enterotoxin B (SEB) from Staphylococcus aureus could be considered as a therapeutic agent to eliminate cancer cells. SEB can activate immune response, which furthermore could induce apoptosis of various cancer cells. This study was proposed to design a SEBsyn coding sequence suitable for Escherichia coli BL21(DE3) expression system for recombinant protein production. This research was conducted by designing the inserted gene that included codon optimization. The obtained synthetic gene was transformed into E. coli BL21(DE3) and the transformants were verified by PCR with T7 promoter and T7 terminator primers. The results showed that the SEBsyn encoding gene could be synthesized and cloned into pET-28a(+). This recombinant plasmid that carried the SEB encoding gene (pET-28a_SEBsyn) was successfully transformed into E. coli BL21(DE3). Amplicon of colony PCR visualized in 2% agarose gel showed that transformants carrying the recombinant plasmid had an inserted gene length about 1,000 bp. In contrast, the amplicon from colonies that carries control plasmid (pET-28a(+) without insert) had a gene length about 300 bp. The protein verification by SDS-PAGE also confirmed that the plasmid carrying the gene insert was successfully overexpressed in E. coli BL21(DE3).

Copyright

© 2022 The Authors. Published by Atlantis Press International B.V.

Open Access

This is an open access article distributed under the CC BY-NC 4.0 license.

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TY  - CONF
AU  - Achmad Rodiansyah
AU  - Marselina Irasonia Tan
AU  - Husna Nugrahapraja
PY  - 2022
DA  - 2022/05/02
TI  - Construction, Cloning, and Overexpression of Staphylococcal Enterotoxin B Gene Synthetic (SEBsyn) in pET-28a(+): Pre-development Bacterial-Toxin Therapy for Cancer
BT  - Proceedings of the 7th International Conference on Biological Science (ICBS 2021)
PB  - Atlantis Press
SP  - 464
EP  - 470
SN  - 2468-5747
UR  - https://doi.org/10.2991/absr.k.220406.065
DO  - 10.2991/absr.k.220406.065
ID  - Rodiansyah2022
ER  -