Difference between revisions of "Part:BBa K5082010"
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===Usage and Biology=== | ===Usage and Biology=== | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
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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
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Illegal PstI site found at 7344 - 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 7332
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Illegal AgeI site found at 6776 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 3752
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