Expression of the major capsid protein of human papillomavirus type 11 in Saccharomyces cerevisae.
The major capsid protein L1 of papillomaviruses expressed recombinantly or in infected cells has the intrinsic ability to form virus-like particles (VLPs) which display conformational epitopes necessary to elicit high-titered, virus-neutralizing antibodies. We have shown previously that the L1 gene of human papillomavirus type 6a (HPV6) can be expressed efficiently in Saccharomyces cerevisae (Sc) as VLPs. However, when we attempted to express the L1 gene cloned from the closely related HPV11 in yeast, few VLPs were observed in crude lysates. The lower expression level of HPV11 L1 protein was found to result from a truncation of the HPV11 L1 mRNA. Since sequence requirements for transcriptional termination in yeast are still unclear, the HPV6 L1 gene was used as the basis for the complete synthetic reconstruction of the entire 1506-bp HPV11 L1 gene. Expression of this HPV6/11 hybrid L1 gene in yeast resulted in predominantly full-length L1 mRNA and a > 7-fold increased level of production of HPV11 VLPs compared to that expressed by the wild-type HPV11 L1 gene. The VLPs were shown to display the conformational epitopes important to elicit virus-neutralizing antibodies.
Neeper
M P
MP
Department of Virus and Cell Biology, Merck Research Laboratories, West Point, PA 19486, USA.
Hofmann
K J
KJ
Jansen
K U
KU
D001483:Base Sequence; D036022:Capsid Proteins; D003001:Cloning, Molecular; D004279:DNA, Viral; D018985:Epitopes, B-Lymphocyte / Q000032:analysis / Q000235:genetics; D005814:Genes, Viral; D008969:Molecular Sequence Data; D009856:Oncogene Proteins, Viral / Q000235:genetics* / Q000276:immunology / Q000648:ultrastructure; D027383:Papillomaviridae / Q000235:genetics* / Q000276:immunology; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000648:ultrastructure; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D014764:Viral Proteins
D036022:Capsid Proteins; D004279:DNA, Viral; D018985:Epitopes, B-Lymphocyte; C094776:L1 protein, Human papillomavirus type 11; D009856:Oncogene Proteins, Viral; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C091560:L1 protein, Human papillomavirus type 6