Perbandingan Pengaruh Agen Plasmid Curing Acriflavine dan Acridine Orange (AO) terhadap Laju Plasmid Curing dengan Kuantifikasi Fluoresensi Ekspresi Protein GFP pada Bakteri Escherichia Coli PRSET-emGFP
DOI:
https://doi.org/10.51967/tanesa.v23i2.1041Keywords:
Plasmid, Plasmid Curing, Acridine Orange, Acriflavine, GFP, E. coli PRSET-emGFP.Abstract
Plasmid curing is an activity to remove plasmids in a bacterium. Plasmid itself is a structure that can carry genes that are beneficial for bacteria, such as making the bacteria resistant to antibiotics. Acridine Orange (AO) and Acriflavine are intercalating agents that function to break down the super helical shape of plasmid DNA and then form circular or open linear plasmid DNA which is used to carry out Plasmid Curing in this study and then compare its effect on the rate of plasmid curing by fluorescence quantification of GFP protein expression. on Escherichia coli PRSET-emGFP bacteria. The results obtained from this experiment were the rate of plasmid loss per generation obtained by measuring the activity of the GFP reporter on the recombinant E. coli model given the curing agent Acridine Orange was 9.52% for the AO 50 treatment, 44.2% for the AO 100, 10.4% for the AO 50 treatment. Akriflavina 250, and 0.836% for Acriflavine 125. Therefore, it can be concluded that the Acridine Orange treatment was more effective than the Acriflavine treatment against the pRSET-EmGFP recombinant E. coli curing plasmid.
References
Biologics International Corp. 2018. IPTG Induction Theory. https://www.biologicscorp.com/blog/iptg-induction-protein-expression: Diakses pada 12/05/2022 pk. 15:02 WIB
Brejc, K., Sixma, T. K., Kitts, P. A., Kain, S. R., Tsien, R. Y., Ormö, M., & Remington, S. J. (1997). Structural basis for dual excitation and photoisomerization of the Aequorea victoria green fluorescent protein. Proceedings of the National Academy of Sciences of the United States of America, 94(6), 2306–2311.
Byvaltsev, Vadim A.; Bardonova, Liudmila A.; Onaka, Naomi R.; Polkin, Roman A.; Ochkal, Sergey V.; Shepelev, Valerij V.; Aliyev, Marat A.; Potapov, Alexander A. (2019). Acridine Orange: A Review of Novel Applications for Surgical Cancer Imaging and Therapy. Frontiers in Oncology, 9(), 925–.
Couturier, M. A. R. T. I. N. E., Bex, F., Bergquist, P. L., & Maas, W. K. (1988). Identification and classification of bacterial plasmids. Microbiological reviews, 52(3), 375-395.
Das, T., Chakraborty, B., & Ray, R. (2017). In-Vitro Study on Potential Efficacy of Acriflavine in the Prevention of Conjugational Transfer of Drug Resistance in Bacteria. J. Evol. Med. Dent. Sci, 6, 5540-5547
Invitrogen. (2010). pRSET/CFP, pRSET/EmGFP, pRSET/BFP Vectors User Manual. Available from https://assets.thermofisher.com/TFS-Assets/LSG/manuals/prsetfpvectors_man.pdf
Kowalska-Krochmal, B., & Dudek-Wicher, R. (2021). The Minimum Inhibitory Concentration of Antibiotics: Methods, Interpretation, Clinical Relevance. Pathogens (Basel, Switzerland), 10(2), 165. https://doi.org/10.3390/pathogens10020165.
MacWilliams MP, Liao MK. 2006. Luria Broth (LB) and Luria Agar (LA) Media and Their Uses Protocol. American Society for Microbiology. www.asmscience.org: Diakses pada 12/05/2022 pk. 0:02 WIB
Mandelstam, J., & McQuillen, K. (1968). Biochemistry of bacterial growth. Biochemistry of bacterial growth. Parker N. 2012. Microbiology. Openstax: p.568-570
Persinoti, G. F., Peres, N. T. A., Jacob, T. R., Rossi, A., Vencio, R. Z. dan Martinez-Rossi, N. M. 2014. RNA-sequencing analysis of Trichophyton rubrum Transcriptome in Response to Sublethal Doses of Acriflavin. BioMed Central. 15(7).
Rozwandowicz, M., Brouwer, M. S. M., Fischer, J., Wagenaar, J. A., Gongalez-Zorn, B., Guerra, B., Mevius, D. J., Hordijk, J. (2018). Plasmids carrying antimicrobial resistance genes in Enterobacteriaceae, Journal of Antimicrobial Chemotherapy, 73 (5), 1121–1137.
Trevors, J. T. (1986). Plasmid curing in bacteria. FEMS microbiology reviews, 1(3-4), 149-157.
Villagra, Nicolás & Hidalgo, Alejandro & Santiviago, Carlos & Saavedra, Claudia & Mora-Longa, Guido. (2008). SmvA, and not AcrB, is the major efflux pump for acriflavine and related compounds in Salmonella enterica serovar Typhimurium. The Journal of antimicrobial chemotherapy. 62. 1273-6. 10.1093/jac/dkn407.
Yektaeian, N., Mehrabani, D., Sepaskhah, M., Zare, S., Jamhiri, I., Hatam, G. (2019). "Lipophilic tracer Dil and fluorescence labeling of acridine orange used for Leishmania major tracing in the fibroblast cells". Heliyon. 5 (12): e03073.
Downloads
Published
How to Cite
Issue
Section
License
The copyright of this article is transferred to Buletin Poltanesa and Politeknik Pertanian Negeri Samarinda, when the article is accepted for publication. the authors transfer all and all rights into and to paper including but not limited to all copyrights in the Buletin Poltanesa. The author represents and warrants that the original is the original and that he/she is the author of this paper unless the material is clearly identified as the original source, with notification of the permission of the copyright owner if necessary.
A Copyright permission is obtained for material published elsewhere and who require permission for this reproduction. Furthermore, I / We hereby transfer the unlimited publication rights of the above paper to Poltanesa. Copyright transfer includes exclusive rights to reproduce and distribute articles, including reprints, translations, photographic reproductions, microforms, electronic forms (offline, online), or other similar reproductions.
The author's mark is appropriate for and accepts responsibility for releasing this material on behalf of any and all coauthor. This Agreement shall be signed by at least one author who has obtained the consent of the co-author (s) if applicable. After the submission of this agreement is signed by the author concerned, the amendment of the author or in the order of the author listed shall not be accepted.
