PHASE 1 :
- Amplification/isolation of the gene of interest out of a customer-supplied vector and subcloning it into a transfer vector with Histag.
- Sequence confirmation of insert.
- Transformation of E. coli with baculoviral DNA and gene of interest..
- Culture of E. coli and purification using mini-prep to produce bacmid (recombinant viral DNA).
- Transfection of sf9 insect cells with bacmid.
- Plaque purification and viral titer assay.
- Small-scale culture to test expression level.
- Estimated time: 4-6 weeks
PHASE 2 :
- Virus stock preparation.
- Large-scale culture and infection of insect cells for protein expression..
- Purification from 1 liter of infected insect cell using affinity column of Ni-NTA or other column.
- Estimated time: 2-4 weeks
DELIVERABLES
5 µg transfer vector, bacmid, recombinant virus,protein, COA.