Expression of simian virus 40 T antigen in insect cells using a baculovirus expression vector.
Simian virus 40 (SV40) large T and small t antigens were synthesized in insect cells using the baculovirus Autographa californica as an expression vector. A recombinant virus containing a genomic copy of the SV40 early region expressed high levels of small t antigen but only low levels of large T antigen. However, very high levels of T antigen synthesis were observed when viruses were constructed with a cDNA copy of the large T antigen mRNA. Insect cells were capable of modifying T antigen by phosphorylation, palmitylation, glycosylation, and oligomerization. Functional assays demonstrated that the origin-specific DNA binding, ATPase, and helicase activities of insect cell-derived T antigen were comparable to T antigen synthesized in mammalian cells. Use of the baculovirus vector system to produce T antigen should facilitate future investigations requiring large quantities of T antigen.
Lanford
R E
RE
Department of Virology and Immunology, Southwest Foundation for Biomedical Research, San Antonio, Texas 78284.
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D001345:Autoradiography; D001483:Base Sequence; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006031:Glycosylation; D007304:Insect Viruses / Q000235:genetics*; D008969:Molecular Sequence Data; D009036:Moths; D010766:Phosphorylation; D010957:Plasmids; D011233:Precipitin Tests; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
D000952:Antigens, Polyomavirus Transforming; D011994:Recombinant Proteins