Question

7) Your goal is to create a bacterial strain that produces a viral envelope protein to be used as an antigen for immunization of humans against this virus.

a) List the information/materials you would need to start this process

b) Taking your list of materials, sketch the steps needed to create the specific bacterial strain

Answer 1

Aim: To create a bacterial strain that produces viral envelope protein

For this, we have to clone the gene of interest (GOI = Viral envelop protein-encoding gene) into a cloning vector followed by subcloning into an expression vector

Material required:
1. Viral template DNA
2. Primers specific to GOI
3. Taq polymerase and other components required for PCR
4. Cloning and expression vectors
5. Restriction enzymes and DNA ligase
6. Agar, antibiotic solutions and other required culturing materials
7. Competent cells

Procedure:
1. Identification of GOI
2. PCR of the GOI with specific primers
3. Restriction digestion of the PCR and a suitable cloning vector
4. Ligation of the vector and insert
5. Transformation into competent cells
6. Plating on selection plate
7. Screening for positive clones
8. Confirmation of positive clones by restriction digestion and sequencing
9. Sub-cloning into a suitable expression vector
10. Induction of protein expression