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Table 2 Comparison of multi-gene expression systems available for the BEVS

From: GoldenBac: a simple, highly efficient, and widely applicable system for construction of multi-gene expression vectors for use with the baculovirus expression vector system

 MultiBac/OmniBacLIC-based MacroBacUSER-based systembiGBacGoldenBac
Number of genesup to 4 with Cre-lox, more possible via restriction/ligationup to 8, more possible with additional cloning roundsup to 16
more possible with additional cloning rounds
up to 25up to 15
Expression of intermediatesno, only those in or fused to acceptorsyesyesyesyes
EfficiencyVaries; triple assembly via Cre-lox has very low efficiency2 colonies to be screened at each step10 colonies to be tested for assembly of dual cassettes into multi-gene constructs6–12 colonies to be tested in first assembly step2 colonies tested for assembly of up to 12 genes
Construct verificationmany possible variants with possibility of Tn7 element duplicationonly 1 correct version at each steponly 1 correct version at each steponly 1 correct version at each steponly 1 correct version
Sequence requirement2–4, none
4+, no BstXI sites
no PmeI, SwaI sitesnone2–5, none
5+, no PmeI sites
no BsaI sites
Number of assembly steps2–4 genes, 1 step
4+, 2 steps
2 genes, 1 step,
4 genes, 2 steps, 8 genes, 3 steps
2–4 genes, 1 step
4+, 2 steps and 2 bacmid integrations
2–5 genes, 1 step
5+, 2 steps
1 step
Method of Baculovirus generationTn7 transposase or homologous recombinationTn7 transposase onlyTn7 transposase
only
Tn7 transposase onlyTn7 transposase or homologous recombination
  1. Information on cloning systems is taken from: MultiBac/OmniBac [17, 27], USER [6], biGBac [2], MacroBac [7]