Proceedings of the 7th International Conference on Biological Science (ICBS 2021)

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

Authors
Achmad Rodiansyah1, Marselina Irasonia Tan1, *, Husna Nugrahapraja1
1Institut Teknologi Bandung, School of Life Sciences and Technology, Department of Biotechnology, Labtek XI, 10 Ganesha street, Bandung 40132, Indonesia
*Corresponding author. Email: marsel@sith.itb.ac.id
Corresponding Author
Marselina Irasonia Tan
Available Online 2 May 2022.
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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Volume Title
Proceedings of the 7th International Conference on Biological Science (ICBS 2021)
Series
Advances in Biological Sciences Research
Publication Date
2 May 2022
ISBN
10.2991/absr.k.220406.065
ISSN
2468-5747
DOI
10.2991/absr.k.220406.065How to use a DOI?
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.

Cite this article

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  -