Early Detection of Productive Baculovirus DNA Transfection

Early detection of productive baculovirus DNA transfection.

The most widely studied baculovirus, the Autographa californica M nucleopolyhedrovirus (AcMNPV), is a large, enveloped DNA virus that infects lepidopteran larvae (8). This virus is commonly used as a eukaryotic vector to express heterologous genes in cultured insect cells and insect larvae (1,3,15). Baculovirus expression systems offer several advantages over other expression systems, among whi...

Transfection of insect cell in suspension for efficient baculovirus generation

Baculovirus (BV) mediated insect cell expression system utilizes transfection as a first step to introduce recombinant baculovirus DNA into insect cells. Many labs are still relying on the conventional liposome based transfection method in adherent culture. Here we describe a more efficient method that can replace the existing method. This method is economical and does not require any special a...

Replication of baculovirus DNA.

The Baculoviridae is a diverse family of pathogens that are infectious for arthropods and are characterized by a complex replication cycle that culminates in the occlusion of virions in a crystalline protein matrix (Blissard & Rohrmann, 1990). Over 400 lepidopteran species serve as hosts for these viruses, with a single virus isolate usually restricted to one or a few related species. In additi...

Neutral Liposomes and DNA Transfection

Glycerol enhancement of ligand-polylysine/DNA transfection.

Primary human fibroblasts and a series of cell lines (A549, BNL CL.2, H225, NIH 3T3 and Rat-1) are efficiently transfected by using positively charged complexes of plasmid DNA and transferrin-polylysine or polylysine in the presence of glycerol (1 molar to 1.8 molar, depending on the cell type). An increase in gene expression of up to several-hundredfold (compared to complexes without glycerol)...