Introduction of sequences encoding functional human adenosine deaminase into mouse cells using a retroviral shuttle system.
A retroviral packaging system was used to generate a murine virus carrying sequences encoding human adenosine deaminase (ADA). To this end, human ADA cDNA was inserted into the retroviral shuttle vector pZIP-NeoSV(X)1. This vector provides all of the cis-acting sequences necessary for the efficient packaging and transmission of the viral genome as well as a selectable gene for G418 resistance. Transfection of this recombinant plasmid into cells that provide essential virus products (psi-2 cells) yielded cell lines that stably produced virions carrying the coding sequence of human ADA. We have used these virions to infect NIH3T3 cells, which after 48 h synthesized catalytically active human ADA. Furthermore, G418-resistant cell lines were obtained from the virus-infected NIH3T3 cells that stably produced the human ADA enzyme.
D000243:Adenosine Deaminase / Q000235:genetics*; D000818:Animals; D002460:Cell Line; D005796:Genes; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D009700:Nucleoside Deaminases / Q000235:genetics*; D012190:Retroviridae / Q000235:genetics; D014170:Transformation, Genetic
D009700:Nucleoside Deaminases; D000243:Adenosine Deaminase