The use of baculovirus as a cloning and expression vector involves following steps :
(i) desired gene is cloned in a transfer vector under the control of polyhedrin promoter;
(ii) chimeric transfer vector is cotransfected with wild type virus into the host cell;
(iii) polyhedrin gene of wild type is replaced by cloned gene through recombination;
(iv) recombinant virus is plaque purified, since they will have no polyhedra occlusion bodies;
(v) the selected recombinant virus is multiplied and used for expression of heterologus protein.
Construction of baculovirus expression vectors
|