A question we are asked from time to time is to identify baculovirus transfer vectors compatible with our flashBAC™ system. We obviously nominate our own extensive range that is available but frequently someone has an old plasmid from years ago that they want to reuse. Originally they may have paired it with a linear DNA system such as BaculoGOLD or BacPAK6 or even wild type DNA where recombinant virus selection was effected by identifying polyhedrin-negative plaques. In the early days of the baculovirus expression system, many labs world wide constructed many different plasmid transfer vectors and there was little consistency in nomenclature. Most were based on the polyhedrin gene promoter flanked by sequences homologous to the baculovirus genome. Cotransfection of insect cells with a transfer vector and virus DNA generated the recombinant virus. This pattern was broken by Verne Luckow’s innovative system where T7 transposition in E.coli was used to introduce the foreign gene into the virus genome. These vectors (e.g. pFastBac) are only compatible with Bac-to- Bac. Unfortunately, you can’t use these with any other virus genome.
The OET flashBAC™ system is compatible with almost any other plasmid transfer vector, including many used for multiple gene expression. To make life easier for those of you wishing to be sure that the plasmid you have will work with flashBAC™, we have put together a table that summarizes all of the appropriate transfer vectors. Currently this includes a list of restriction enzyme sites for the insertion of foreign sequences. In future we hope to insert links to vector sequences too.
If we haven’t listed the plasmid that you have or you are still in doubt, don’t hesitate to get in touch with us at OET Ltd and we will try to help you.