Bacmid DNA can be generated using high-throughput technology. This is useful for rapid and efficient testing of variables like the promoter used for expression, expression tags (FLAG, 10xHis, fluorescent, or other fusion protein) and their position (N- or C-terminal), truncations, mutations, cleavage sites, or even different media for ...

molecular weight recombinant bacmid DNA and transfect the bacmid DNA into insect cells using the ExpiFectamine ™ Sf Transfection Reagent to generate a recombinant baculovirus that can be used for preliminary expression experiments.

The bacmid system exploits two remarkable technologies: the ability of the bacterial transposon Tn7 to specifically integrate adjacent to a defined sequence in bacterial genomes, and the identification of the mini-F plasmid replicon that, when incorporated into the baculovirus genome, allows its coordinated replication in bacteria as an ...

There are specific kits for isolation of large DNA episomes such as BACs that may be good for you. This is the protocol I was using back in the dark ages. B. Isolation of Bacmid: 1. Grow...

Feb 7, 2022 · Linearized bacmid DNA are commercially available but the cost can be prohibitive when a large number of viruses need to be generated. Here we describe a baculovirus expression toolbox (HR-Bac)...

This protocol details the construction of bacmid DNAs where a baculovirus-expression-system-style expression clone is transformed into an E. coli strain containing a baculovirus shuttle vector and transposases on a helper plasmid. Once the transposition is confirmed, bacmid DNA is extracted to be used for propagating recombinant baculovirus.

There are two major strategies to genetically modify baculoviruses. One uses a bacmid-based system which replicates in Escherichia coli using a bacterial origin of replication. The other employs a transfer vector and viral DNA which are co-transfected into insect cells and utilise host enzyme-mediat …