Fluorescent protein expression in Ecoli
- 1 min
The expression of an unknown fluorescent protein in E. coli was confirmed through several molecular techniques. First, I transformed the gene into E. coli using heat shock, followed by plating the cells on antibiotic-containing media to select for those that successfully took up the plasmid. I then performed a miniprep to extract the plasmid DNA from the transformed cells. To verify the presence of the gene, I conducted PCR with specific primers and performed a restriction digest, analyzing the resulting fragments with gel electrophoresis. Sequencing confirmed the correct gene sequence. For protein expression, I induced the transformed E. coli with IPTG and confirmed the expression by observing fluorescence under a microscope. Finally, I used SDS-PAGE and Western blot to validate the size and presence of the fluorescent protein with specific antibodies