Expression of human papillomavirus proteins in yeast Saccharomyces cerevisiae.
The L1 and L2 proteins of human papillomavirus (HPV) types 1, 6, and 16 and the E6 and E7 proteins of HPV 16 were expressed in Saccharomyces cerevisiae. The yeast expressed proteins were readily detected by immune blotting and were generally intact. The HPV 1 L1 and L2 proteins expressed in yeast were indistinguishable from the major and minor capsid proteins purified from HPV 1 virions as judged by gel electrophoresis and immunoblotting. The HPV 6 and HPV 16 L2 proteins and HPV 16 E7 proteins were secreted from yeast by fusion to the yeast pre-pro-alpha-factor leader sequence. Following secretion of the HPV 16 E7 protein a rapid method of purification was developed. The yeast expressed proteins were used as antigen targets to study the human immune response in Western blot assay, ELISA, and immune precipitation. One human serum reacted with intact, but not denatured HPV 16 L2 proteins, suggesting that the yeast expressed proteins will be useful to detect antibodies reactive with conformational epitopes.
Carter
J J
JJ
Fred Hutchinson Cancer Research Center, Seattle, Washington 98104.
Yaegashi
N
N
Jenison
S A
SA
Galloway
D A
DA
D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics*; D003001:Cloning, Molecular; D015870:Gene Expression; D006023:Glycoproteins / Q000235:genetics; D006031:Glycosylation; D006801:Humans; D066298:In Vitro Techniques; D008970:Molecular Weight; D027383:Papillomaviridae / Q000235:genetics* / Q000276:immunology; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics* / Q000276:immunology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D006023:Glycoproteins; D011994:Recombinant Proteins; D014764:Viral Proteins