In vitro analysis of expression vectors for DNA vaccination of horses: the effect of a Kozak sequence.
One of the prerequisite for developing DNA vaccines for horses are vectors that are efficiently expressed in horse cells. We have analysed the ectopic expression of the human serum albumin gene in primary horse cells from different tissues. The vectors used are of pcDNA and pUC origin and include the cytomegalovirus (CMV) promoter. The pUC vectors contain CMV intron A whereas the pcDNA vectors do not. Insertion of intron A diminished the expression from the pcDNA vectors whereas insertion of a Kozak sequence upstream of the gene in two types of pUC vectors increased significantly the in vitro expression in primary horse cells derived from skin, lung, duodenum and kidney. We report for the first time the significance of full consensus Kozak sequences for protein expression in horse cells in vitro.
Olafsdóttir
Gudbjörg
G
Institute for Experimental Pathology, University of Iceland, Keldur, Reykjavík, Iceland. gudbjol@hi.is
Svansson
Vilhjálmur
V
Ingvarsson
Sigurdur
S
Marti
Eliane
E
Torsteinsdóttir
Sigurbjörg
S
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western / Q000662:veterinary; D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology; D006736:Horses / Q000276:immunology*; D000077190:Interferon alpha-2; D016898:Interferon-alpha / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins; D012709:Serum Albumin / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection / Q000379:methods / Q000662:veterinary; D019444:Vaccines, DNA / Q000235:genetics* / Q000276:immunology
D000077190:Interferon alpha-2; D016898:Interferon-alpha; D011994:Recombinant Proteins; D012709:Serum Albumin; D019444:Vaccines, DNA