Difference between revisions of "Part:BBa K5082008"

Latest revision as of 06:47, 15 September 2024

pCMV-EGFP 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.

pCMV-EGFP, also known as pCMV-GFP, is a mammalian expression vector of the GFP reporter gene with a CMV promoter [3]. The pCMV-EGFP plasmid can be amplified by DH5α bacterial cells. Meanwhile, it carries an ampicillin resistance gene for selection. The plasmid map for pCMV-EGFP is shown in Figure 1. Previously, this plasmid has been used in scientific research [4].

                      
                      Figure 1. pCMV-EGFP plasmid map.

Design The above characteristics have made pCMV-EGFP 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 pCMV-EGFP 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. pCMV-EGFP bacterial cell culture.

Reference

References

[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/11153/

[4] Matsuda, Takahiko, and Constance L Cepko. “Electroporation and RNA interference in the rodent retina in vivo and in vitro.” Proceedings of the National Academy of Sciences of the United States of America vol. 101,1 (2004): 16-22. doi:10.1073/pnas.2235688100

Sequence and Features