Difference between revisions of "Part:BBa K5082010"

Latest revision as of 06:52, 15 September 2024

pMIR plasmid vector

Usage and Biology

Plasmids are double stranded circular DNA molecules, which carry non-essential genes, found in bacterial cells [1]. Plasmids are multifunction, easy to edit, and can be amplified quickly by bacterial cellular machinery [2]. These advantages have made plasmids a popular tool in genetic research.

pMIR is a mammalian expression vector of the Luciferase reporter gene with a CMV promoter [3]. The pMIR plasmid can be amplified by DH5α bacterial cells. Meanwhile, it carries an ampicillin resistance gene for selection. The plasmid map for pMIR is shown in Figure 1.

                       
                             Figure 1. pMIR plasmid map.

Design

The above characteristics have made pMIR an ideal plasmid for our experiment. In our experiment, we fused an HSU structure downstream the Luciferase gene to visualize G3BP1 concentration and thereby diagnosed gastric cancer. Meanwhile, we also transfected empty plasmid backbones and plasmids with MUT regions downstream GFP into stomach cell lines to serve as control groups.

Characterization

The pMIR plasmid carries an ampicillin resistance gene. Therefore, we cultured bacterial cells containing the plasmid on solid medium with ampicillin. As shown in Figure 2, the bacteria were successfully cultured, proving that the plasmids have been successfully transformed.

                    
                          Figure 2. pMIR bacterial cell culture.

Reference

[1] Carattoli, Alessandra. “Plasmids in Gram Negatives: Molecular Typing of Resistance Plasmids.” International Journal of Medical Microbiology, vol. 301, no. 8, Dec. 2011, pp. 654–658

[2] Patron, N.J. Synthetic Biology and Gene Cloning. Elsevier EBooks, Elsevier BV, 1 Jan. 2017, pp. 112–117

[3] “Addgene: PCMV-GFP.” Www.addgene.org, www.addgene.org/vector-database/3582/.

Sequence and Features