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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/OmniBac

LIC-based MacroBac

USER-based system

biGBac

GoldenBac

Number of genes

up to 4 with Cre-lox, more possible via restriction/ligation

up to 8, more possible with additional cloning rounds

up to 16

more possible with additional cloning rounds

up to 25

up to 15

Expression of intermediates

no, only those in or fused to acceptors

yes

yes

yes

yes

Efficiency

Varies; triple assembly via Cre-lox has very low efficiency

2 colonies to be screened at each step

10 colonies to be tested for assembly of dual cassettes into multi-gene constructs

6–12 colonies to be tested in first assembly step

2 colonies tested for assembly of up to 12 genes

Construct verification

many possible variants with possibility of Tn7 element duplication

only 1 correct version at each step

only 1 correct version at each step

only 1 correct version at each step

only 1 correct version

Sequence requirement

2–4, none

4+, no BstXI sites

no PmeI, SwaI sites

none

2–5, none

5+, no PmeI sites

no BsaI sites

Number of assembly steps

2–4 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 generation

Tn7 transposase or homologous recombination

Tn7 transposase only

Tn7 transposase

only

Tn7 transposase only

Tn7 transposase or homologous recombination

  1. Information on cloning systems is taken from: MultiBac/OmniBac [17, 27], USER [6], biGBac [2], MacroBac [7]