Lecture 21: Biotech in use

Question 1

What factors would you take into consideration when choosing a system to produce your recombinant protein of interest?

Answer 1

Can the system provide all the necessary steps eg. glycosylation…

Is this the cheapest most efficient method?

Question 2

Describe the steps required to make a recombinant protein. (GTEHP)

Answer 2

GTEHP (Get The Egg Harry Potter)

Gene Cloning:
Insert the gene encoding the protein of interest into a vector (plasmid).

Transformation:
Introduce the vector into host cells (bacteria, yeast, or mammalian cells).

Protein Expression:
Cultivate the host cells under conditions that induce protein production.

Protein Harvesting:
Isolate the protein from the cells or culture medium.

Purification:
Purify the protein to remove impurities and obtain the final product.

Question 3

In general terms, how do we engineer proteins for biotechnology? (Similar to GTEHP)

Answer 3

1. Identify a need
2. Target selection
3. Design the protein
4. Gene synthesis
5. Expression and purification
6. Characterisation and optimisation
7. Validation and testing

Question 4

What is the general concept of gene therapy?

Answer 4

Gene therapy involves introducing, removing, or altering genetic material within a person’s cells to treat or prevent disease. The general concept includes:

Targeting:
Identifying the faulty or missing gene responsible for a disease.

Delivery:
Using a vector (often a virus) to deliver the correct or modified gene into the patient’s cells.

Correction:
The new or altered gene helps to correct or compensate for the genetic defect, thereby treating or preventing the disease.

The goal is to address the underlying cause of genetic disorders rather than just treating symptoms.

Question 5

How could gene therapy be used to permanently treat type I diabetes?

Answer 5

Problem:
Pancreatic B-cells destroyed
* autoantibodies recognise proteins on beta cell surface
* no insulin produced.

Solution?
Reprogram pancreatic A-cells to behave as B-cell

Question 6

Explain an example of a non-medical application of recombinant DNA technology.

Answer 6

Generating recombinant proteins in plant crops has the potential to solve climate problems and enhance our ability to feed a growing population.

Question 7

A protein requires glycosylation: Describe three key features for a plasmid to produce this protein:

Answer 7

(there are other options than these three…)

The plasmid will need a mammalian promoter (the protein requires glycosylation so will need mammalian cell culture)

It will need an origin of replication to allow a bacterial cell culture to replicate many copies of the vector to be used

An antibiotic resistance gene so that only bacteria that have taken up the vector will survive (selection tool)

Question 8

Describe one way a glycosylated protein produced could be introduced into human cells?

Answer 8

Via gene therapy, using a viral vector to deliver the proteins DNA into cells

OR

using recombinant proteins produced in a eukaryotic system that have been purified and could be injected.