AB Vector - Baculovirus tutorial

  Baculovirus tutorial
  Rules to achieve about as high recombinant protein expression levels using baculoviruses as in E.coli.
 
                                         Overview

  Baculovirus Technology

Plasmid vectors

Baculovirus DNA

Maintenance of insect cells

Requirements to monolayers

Transfection

Baculovirus stock propagation

Protein expression
  Baculovirus Tutorial
    Gently mix 0.1 μg of plasmid DNA
containing a gene of interest, 5 μl of
baculovirus DNA and H20 up to 50 μl.
     Add dropwise 50 μl
     of 10% Profectin™
     to the DNA mixture
     from step 1.
          Add dropwise DNA-Profectin™ emulsion from
          step 2 to semi-confluent monolayer of insect
          cells. Incubate for 60-72 h, harvest recombinant
          baculovirus and study protein expression.