It’s lucky week number 7. After seven weeks and seven blogs, I finally plan on connecting some of my work in the lab to my research question 🙂
One quick note: If you haven’t read last week’s blog, I recommend you do in order to understand this week’s. If you want to learn more behind the intricacies of the procedure I carried out this week, click here.
You will recall that last week I mentioned that I would be conducting Restriction Enzyme Digestion on the DNA pET30-6H-Flu-A-NC. However … unfortunately … someone else needed to use that DNA … so I moved onto another one called PCMV/hr1. I digested PCMV/hr1 with the restriction enzymes BgL-II and MluI. The restriction sites (where the enzymes cut DNA) are displayed below:
|Diagram 1: Restriction Site for BgL-II||Diagram 2: Restriction Site for MluI|
These restriction enzymes cut DNA into fragments of different sizes. To separate the fragments of DNA by fragment size, gel electrophoresis is conducted. The results of the gel electrophoresis for PCMV/hr1 are below:
|Diagram 3: DNA Ladder of PCMV/hr1 digested by BgL-II and MluI. The bands that have travelled further away from the top are smaller, while those that have travelled less are larger. More information about this image will be in my final research paper.|
So, for those of you who have read my past six blogs, the lab procedures I have carried out so far include:
- Plasmid Mini-Prep
- Restriction Enzyme Digestion
- Gel Electrophoresis
But what do any of these lab procedures have to do with my initial research question: How can we utilize recombinant DNA technology to halt the ongoing cell division of cancerous cells?
Well, to answer my research question, I plan on using recombinant DNA technology to produce tumor suppressor proteins that can be harvested for medicinal use in chemotherapeutic drugs. Many of the lab procedures I have carried out thus far are used in the process to produce proteins using DNA technology:
- Plasmid Mini-Prep: used to obtain a small plasmid DNA from a bacterial species
- Restriction Enzyme Digestion: obtain specific desired fragments of the obtained DNA (a.k.a. Obtaining the gene of interest)
- Gel Electrophoresis: separates fragments of DNA based on fragment size; enables use to obtain the correct gene of interest based on size
So, what’s left for me to do (I do NOT know if I will be able to conduct all three of these processes in the lab, but I will definitely be doing one of them … stay tuned) :
- DNA ligation: connect gene of interest with another plasmid to be inserted into a bacterial cell
- Cloning: Let the bacteria reproduce, expressing the gene of interest resulting in the production of multiple copies of a specific protein
- Protein Purification: harvest and purify the protein from the bacterial cell
Thus, I now connect my internship and research question. The procedures I have so far carried out in the lab are necessary for the production of a protein using recombinant DNA technology. Therefore, the internship has provided insight into how many of the protein production process using recombinant DNA technology is carried out. Only one question is left to answer: Can the same technology be used to produce tumor suppressor proteins and tumor antibodies? Well, that is where my independent research comes in.
So, I finally have connected it all: my research question, the internship, and independent research. I can’t wait to share all my findings in my final research paper. And that’s a wrap for this week. Thanks for reading, and stay tuned for next week’s blog post! Cheers!