Purification of 3×Myc PKG-Puro-Poly A Gene for Bacterial Transformation

Khairil Pahmi, M. Sidratullah

Abstract


Dysregulation of the oncoprotein c-Myc (Myc) is involved in many types of cancer. Myc is a sequence-specific transcription factor that regulates the transcription of genes involved in the control of cell proliferation and apoptosis through mechanisms that are not well understood. The method of this research is experimental. The experiment result will be described.  Some processes will be done by polymerase chain reaction (PCR) to amplify the gene and then it will be extracted to pure the targeted gene. This research has been succesful to purify 3×Myc PKG-Puro-Poly A gene. c-MYC (hereinafter MYC) is an oncoprotein consisting of 439 amino acids contains a well-characterized C-terminal DNA compound and an N-terminal transactivation domain (TAD). The C-terminal region »100 residues comprises a basic leucine zipper-helix-loop-helix (bHLH-LZ) segment that regulates heterodimerization between MYC and its partner bHLH-LZ MAX mediate in their binding to gene promoters.

Full Text:

PDF

References


Liu X, Tesfai J, Evrard YA, Dent SYR, Martinez E. c-Myc Transformation Domain Recruits the Human STAGA Complex and Requires TRRAP and GCN5 Acetylase Activity for Transcription Activation. J. Bio. Chem. 2003; 278 (22): 20405-20412. https://www.sciencedirect.com

Wang C, Zhang J, Yin J, Gan Y, Xu S, Gu Y and Huang W. Alternative approaches to target Myc for cancer treatment. Signal Transduction and Targeted Therapy. 2021; 6 (117). https://www.nature.com

Ahmadi SE, Rahimi S, Zarandi B, Chegeni R and Safa M. MYC: a multipurpose oncogene with prognostic and therapeutic implications in blood malignancies. J Hematol Oncol. 2021; 14 (121). https://jhoonline.biomedcentral.com

Casey SC, Baylot V, and Felsher DW. The MYC oncogene is a global regulator of the immune response. Blood 2018; 131 (18). https://ashpublications.org/blood

Llombart V, and Mansour MR. Therapeutic targeting of “undruggable” MYC. eBioMedicine 2022;75. www.thelancet.com

de Alboran IM, O’Hagan RC, Ga¨rtner F, Malynn B, Davidson L, Rickert R, Rajewsky K, DePinho RA, Alt FW. Analysis of C-MYC Function in Normal Cells via Conditional Gene-Targeted Mutation. Immunity, 2001; 14: 45–55.

Mundo L, Ambrosio MR, Raimondi F, Del Porro L, Guazzo R, Mancini V, Granai M, Rocca BJ, Lopez C, Bens S, Onyango N, Nyagol J, Abinya N, Navari M, NdedeI, Patel K, Piccaluga PP, Bob R, de Santi MM, Russell RB, Lazzi S, Siebert R, Stein H & Leoncini L. Molecular switch from MYC to MYCN expression in MYC protein negative Burkitt lymphoma cases. Blood Cancer J. 2019; 9:91. https://doi.org/10.1038/s41408-019-0252-2

Levens DL. Reconstructing MYC. 2023 [cited 14 December 2023. Available from : https://genesdev.cshlp.org/content/17/9/1071.full

Stine ZE, Walton ZE, Altman BJ, HsiehAL, and Dang CV. MYC, Metabolism, and Cancer. Cancer Discov. 2015; 5(10) : 1024-39.

Farina A, Faiola F, and Martinez E. Reconstitution of an E box-binding Myc:Max complex with recombinant full-length proteins expressed in Escherichia coli. Protein Expr Purif. 2004; 34(2): 215–222.

Wertz JE, Low KB. Genetics, Microbial (general). Philadelphia : Elsevier; 2017.

Dean, D. Bacterial Transformation: The Heat Shock Method. 2023 [Cited 04 December 2023]. Available from : https://www.jove.com/v/5059/bacterial-transformation-using-heat-shock-and-competent-cells

Peukert K, Staller P, Schneider A, Carmichael G, Hänel F, Eilers M. An alternative pathway for gene regulation by Myc. The EMBO Journal (1997)16:5672-5686. https://doi.org/10.1093/emboj/16.18.5672

Miyamoto C, Chizzonite R, Crowl R, Rupprecht K, Kramer R, Schaber M, Kumar G, Poonian, And Jut G. Molecular cloning and regulated expression of the human c-myc gene in Escherichia coli and Saccharomyces cerevisiae: Comparison of the protein products. Proc. Nati. Acad. Sci. USA 1985; 82: 7232-723.

Dang CV, Dam HV, Buckmire M, and Lee WMF. DNA-Binding Domain of Human c-Myc Produced in Escherichia coli. Mol. and Cell. Bio. 1989; 9 (6): 2477-2486.




DOI: https://doi.org/10.37311/jsscr.v5i3.24069

Refbacks

  • There are currently no refbacks.


Copyright (c) 2024 Journal Syifa Sciences and Clinical Research

EDITORIAL OFFICE OF JOURNAL SYIFA SCIENCES AND CLINICAL RESEARCH
  Department of Pharmacy, Gorontalo State University
Jl. Jenderal Sudirman No.6, Kota Gorontalo, Provinsi Gorontalo 96128, Indonesia
 Email: info.jsscr@ung.ac.id
Google Scholar : JOURNAL SYIFA SCIENCES AND CLINICAL RESEARCH

Creative Commons License

Journal Syifa Sciences and Clinical Research (JSSCR) is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.