B-Galalactosidase Recombinant Aden

Background

Recombinant adenoviruses have enormous potential in both research and therapeutic applications. There are numerous advantages in using adenovirus to introduce genetic material into host cells. The range of permissive host cells is very wide. The virus has been used to infect many types of mammalian cells (both replicative and non-replicative) for high expression of the recombinant protein.

Recombinant Adenoviruses are especially useful for gene transfer and protein expression in cell lines that have low transfection efficiency with liposomes. After entering cells, the virus remains epichromosomal (that is, it does not integrate into the host’s chromosome, so it does not turn the host’s genes on or off). Recently, recombinant adenoviruses have been used to deliver RNAi to cells.

Safety Consideration

Remember that you will be working with samples that contain infectious viruses. Follow the recommendations NIH guidelines for all materials containing BSL-2 organisms. Always wear gloves, use filtered tips and work under a biosafety hood.

Methods

The proper amount of virus used to infect cells is critical to the outcome of your experiments. If not enough virus is used, it will not give 100% infection. If too much virus is used, it will cause cytotoxicity or other unwanted effects. The number of adenovirus cell surface receptors varies greatly between different cell types, therefore the optimal concentration differs dramatically between cell types. TO a range of 10-200 MOI (multiplicity of infection) is used for most cell lines, but up to 1000 MOI can be used for lymphoid cell lines.

Traditionally, infectivity particles are measured in culture by a plaque-forming unit (PFU) assay that Scores the number of viral plaques based on dilution. In contrast to the 10-day infection of a Classic Plaque Assay, Cell Biolabs QuickTiter ™ Adenovirus Titer Immunoassay Kit (Cat. # VPK-109) it only requires a 2-day infection and there is no agar overlay step.

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