Table 3.
Comparisons among MultiBac, biGBac and SmartBac systems.
| MultiBac | biGBac | SmartBac | ||
|---|---|---|---|---|
| Expression Strategy | Multi-GEC | Multi-GEC | Polyprotein & Multi-GEC | |
| Vectors | Acceptors (pACEBac1, pACEBac2) Donors (pIDS, pIDK, pIDS) |
pLIB, pBIG1, pBIG2 | Acceptors (4V1G, 4V1R, 5V1TG, 5V1TR) Donors (4V2G, 4V2R) |
|
| Vectors that can produce recombinant baculoviruses | Only acceptors | pLIB, pBIG1, pBIG2 | Only acceptors | |
| Molecular cloning | Construction of junior plasmid containing one GEC | Use the acceptors and donors | Use only the pLIB | This step is not needed. |
| Construction of vectors containing multiple genes | Construct large acceptors and large donors using the multiplication module consisting of homing endonuclease and BstXI | Construct pBIG1 vectors by linker-optimized Gibson assembly | Construct large acceptors and large donors by overlapping PCR and Gibson assembly | |
| Method to screen positive clones | None | None | Blue-White screening | |
| Construction of vectors containing more genes | Recombine large acceptor and large donor by Cre-LoxP combination | Construct pBIG2 vector by linker-optimized Gibson assembly from pBIG1 vectors | Recombine large acceptor and large donor by Cre-LoxP combination | |
| Whether the vector supports expression monitoring | No | No | Yes (using 4V1G and 4V1R) | |
| Whether the vector supports transfection and virus amplification monitoring | No | No | Yes | |
| Whether the vector supports monitoring co-infection of two kinds of viruses | No | No | Yes | |
| Reported largest protein complexes expressed | Dynein complex (6 cDNAs, 1.4 MDa) | APC/C related complex (17 cDNAs, 17 subunits, 1.2MDa) | Dynactin complex (11 cDNAs, 23 subunits, 1.2MDa) | |