Abstract
Generation of stable cell lines is a widely used technique for continuous recombinant protein production. Advantages of the constitutive stable over the transient protein expression are uniformity of the expression across cell populations as well as high quantity and consistency of the protein yields. This data describe step-by-step procedure for the production of glycoprotein without a transmembrane domain (GDTM) of bovine ephemeral fever virus (BEFV) by mammalian stable cells. LentiX-293T cells were transfected with four plasmid constructs to generate a recombinant lentivirus. Subsequently, 293T cells were transduced by the recombinant virus and the polyclonal stable cell pools were then selected by puromycin. Next, limiting dilution was performed from each cell pool to isolate the monoclonal stable cells expressing GDTM protein. Western blot analysis showed that all monoclonal cell clones could stably express GDTM protein. The data confirms that the stable 293T cell line expressing the secretory GDTM protein is an attractive platform for antigen production. (c) 2019 The Author(s). Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
