Several unique features of the Baculovirus System make this virus very appealing for preventive or therapeutic gene transfer. These include:

• The ability to generate large amounts of recombinant protein
• Expression of genes from bacteria, viruses, plants, and mammals at levels from 1-500mg/L
• Several post-translational modifications possible; phosphorylation, glycosylation, and acylation

By pairing this technology with our expertise in virus packaging, abm offers custom Baculovirus subcloning and virus production, allowing high-level gene expression in insect cells.

Custom Baculovirus Plasmid Cloning

Custom Cloning Service

Custom Baculovirus Production

Baculovirus Controls

1. C173 or C174: These services must be ordered with a Custom Baculovirus Production service, either C148 or C149.
2. C098 or C129: These services must be ordered with a Custom Baculovirus Production service, either C148 or C149, and a Custom Baculovirus Cloning service, either C173 or C174.
3. ***Cloning of genes over 3kb are subject to an additional charge of $100.00/kb (C100). Please contact technical@abmgood.com for more information on available cloning vectors and restriction enzyme sites to qualify for the compatible insert service.
4. For customer-supplied vectors, the customer must indicate prior to the service if any reporter expression will be expected or not. The expected reporter expression, e.g. GFP, should not require induction of expression. Due to differences in excitation/emission wavelengths for different fluorophores, we may not be able to provide infection images for fluorescent reporter expression other than standard GFP/RFP/mCherry.

An overview of abm’s custom Baculovirus service:

Baculovirus is a helper-independent viral system that is used to express heterologous genes. Because of their ability to efficiently infect a wide range of hosts, and their capacity to generate high yields of gene targets, the Baculovirus Expression Vector System (BEVS) has shown considerable promise to establish long-term gene expression in most cells. Polyhedrin and p10 are considered non-essential genes when Baculovirus is in culture, allowing the coding sequences to be replaced with a sequence for the target protein. Compared to bacterial expression systems, the post-translational processing and folding of recombinant proteins produced in insect cells more closely resembles mammalian processes, and the yields of functional protein are often much greater.

Step 1: Gene Synthesis and/or Subcloning

• Strategy design and codon optimization
• Gene synthesis or amplification/isolation of the gene of interest from a customer-supplied vector and subcloning it into a transfer vector with His tag
• Sequence confirmation of insert
• Estimated time: 2 weeks

Step 2: Virus Generation and Expression Evaluation

• Transformation of gene of interest into E. coli with baculoviral DNA
• Culture of E. coli and purification using mini-prep to produce an expression bacmid (recombinant viral DNA)
• Transfection of Sf9 insect cells with expression bacmid
• Plaque purification and viral titer assay
• Small-scale culture to test expression level (His-tagged proteins only)
• Estimated time: 4-6 weeks

Optional: Protein Purification

• Virus stock preparation and amplification
• Large-scale culture and infection of insect cells for protein expression
• Purification from 1 litre of infected insect cell using affinity column of Ni-NTA or other column
• Estimated time: 4-5 weeks

Site Directed Mutagenesis service