Introduction
write a literature review about prmt5 role, function and regulation
Body paragraphs
Identify the open reading frame by highlighting in bold red text the start and stop codons on the FASTA cDNA sequence included in the appendix, and calculate the predicted molecular weight of the protein produced by the open reading frame. (1 point)
Propose an experiment that can allow you to identify its interacting partner proteins (a summary scheme of your experiment can be included in appendix B after the FASTA cDNA nucleotide sequence), and include positive and negative controls in your experimental design. (1 point)
Generate a restriction map of your cDNA, and propose a cloning strategy of your cDNA (including the open reading frame) into a eukaryotic expression vector (a summary scheme of your cloning strategy along with a circular map of the cloning vector should be included in appendix C). (1 point)
You wish to investigate the biological function of your gene of interest in a cancer cell line, where you discover that expression of your gene of interest is dysregulated. You check its mRNA and protein expression and you are able to confirm that misexpression of your gene of interest stems from the fact that it is post-transcriptional. Therefore, you decide to determine if there are miRNAs and or tsRNAs involved in its regulation.
A. Using the miRNA search engine (https://mirbase.org/), provide the list of potential miRNAs involved in the regulation of your gene of interest (the miRNA list should be included in appendix D), and highlight in bold green text the binding sites of the strongest miRNA binding sites on the cDNA provided in appendix A. In addition, provide a short overview of what miRNA are (1/2 – 1 page). (1 point)
B. Using the tsRNA search engine (https://rna.sysu.edu.cn/tsRFun/), provide the list of potential tsRNA involved in the regulation of your gene of interest (the tsRNA list should be included in appendix E), and highlight in bold blue text their binding sites on the cDNA provided in appendix A. In addition, provide a short overview of what tsRNA are (1/2 – 1 page).
conclusion
In the conclusion of your report, assume that the gene you selected is dysregulated – Discuss the best possible experiment you can use to
Appendices A-E: Should include in the order shown bleow (1 point)
Appendix A (cDNA sequence with highlighted relevant information as specified in the instructions).
Appendix B (Summary scheme of experiment about interacting proteins).
Appendix C (Summary scheme of cloning strategy along with a circular map of the cloning vector).
Appendix D (List of miRNAs).
Appendix E (List of tsRNAs).
The report should be written with Arial font size: 11, spacing: 1.5. No pictures are allowed in the report 7 pages minimum .
Please do not forget to number your references and to cite the references in your report.
correct expression of your gene of interest in a cancer cell line.
