Assignment 2 BS934 Gene Technology and Synthetic Biology 2022/2023
|Electronic Deadline:||15-NOV-2022 16:00:00|
Design and validation of an expression vector
In the previous assignment you generated a PCR product of your gene of interest. In this assignment, you will describe how you insert the PCR product into a mammalian or bacterial expression vector and then validate the vector. For this, you should generate a 5 minute/ 5 slide presentation (not more, however, it might be shorter, requiring fewer slides!). Please, make this presentation individually yourself.
There are three ways you can generate the presentation:
- Make a Powerpoint presentation and insert audio recordings in each slide (in Powerpoint. To do this, you open ‘insert’, then ‘audio’ the ‘record audio’, then you insert the audio in the slide. You can save this as a Powerpoint file (e.g., .pptx) or Powerpoint show (.ppsx). The upload the presentation onto FASER. Please, be aware that you can only upload < 50 Mb files onto FASER.
- Make a Powerpoint presentation, record yourself using ZOOM presenting this (using share presentation), note down the shareable link to the presentation on a word document (this will be sent to your email address a few hours after you recorded onto ZOOM) and then upload this onto FASER. Please, test that the shareable link will actually lead to your presentation. I do not think FASER has enough memory to accommodate large mp4 files (and they get large), so do not try to upload these.
When you do this (and also (3) or (4), please, upload your powerpoint presentation onto FASER, too, just in case.
- Alternatively, you can set up a YOUTUBE account (takes a few minutes and does not cost anything), upload the mp4 file to YOUTUBE and share the link of your YOUTUBE presentation on FASER (e.g. on a WORD doc). Again, test and make sure the link leads to your presentation, please. Setting up a YOUTUBE presentation is easy and would be my preferred route, because there have been issues with accessing ZOOM links.
- You can also have a combination of (1-3), which is what I did for my YOUTUBE presentations, I recorded onto my Powerpoint presentations and then showed that in a ZOOM recording to generate a mp4 file then uploaded this onto YOUTUBE. By inserting the recording into each slide, you will have more control, e.g., regarding timing or if you wish to rephrase something.
It would be nice if you can show yourself speaking during the presentation, but this is NOT essential.
Now to the content:
The challenge will be to have the right balance between providing sufficient detail and being succinct enough to keep this into 5 minutes. Keep in mind that we have sometimes to provide complex scientific research presentations in 3 minutes, so 5 minutes is a lot of time.
Here are the points that you should try to cover:
- Briefly outline the aim of your project (as an example: “I which to express protein X in mammalian cells to test if expression of the protein will affect the cell cycle” or “I wish to express protein Y as a GFP fusion protein to determine its localisation in a mammalian cell, is it nuclear or cytoplasmic”). Some brief outline about the function of your gene of interest would be good, in line of what you did for assignment 1.
- Outline your strategy to insert your insert into an expression vector (e.g., bacterial or mammalian). Justify why you decided on this particular strategy. Use schemes to illustrate your strategy. Try to be as concrete as possible (e.g., “I will use pET-DEST42 which utilizes the T7/lac promoter for regulated, high-level expression in E. coli”).
- Outline how you transform bacteria and then select for those that contain the correct vector. I will describe techniques to transform bacteria and mammalian cells in my next lectures.
- Outline how you would ensure that you generated the correct vector, e.g. sequencing strategy or any other validation strategy (for example, you might express your protein with a tag, transform cells and perform an immunoblot for your protein of interest).
- Describe the final step of this project, e.g., how you would transform the vector into a mammalian cell of your choice or coli for bacterial expression.
List references or web sites where appropriate in a way that they can be identified.
I attach the marking criteria below, they are the generic marking criteria for presentations.
What I will look for is how much thought you put into the presentation, how concrete it is.
So, please, no general discourse is required (e.g., on the ‘power of molecular biology’ or how sequencing works in detail). I will also mark the design of the slides (make them attractive and do not cram in text) and the overall quality of presentation. So, please, speak clearly, not too fast, not too slow and try to be engaging.
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