The p1 protein of the yeast transposon Ty1 can be used for the construction of bi-functional virus-like particles.
Virus-like particles (VLPs) containing heterologous proteins are often used as vaccines. Two approaches for the construction of bi-functional VLPs using hybrid protein pl-380 of the TY1 transposon of Saccharomyces yeast are described. We have shown that both C- and N-termini of p1-380 can be used for the expression of heterologous peptides. Peptides from A. Fumigatus Asp f 2, expressed at the C- and/or N-termini of p1-380, did not interfere with VLP self-assembling, were accessible for antibodies and hence were exposed at the VLP surface. Another way to obtain bivalent VLPs is the formation of mixed particles, which co-express two hybrid pl proteins with different heterologous protein fragments at the C-terminus. To do it the yeast cells were transfected with a mixture of two recombinant DNA coding Asp f 2 peptide and green fluorescent protein (Gfp). We have shown that both Asp f 2 peptide and Gfp are expressed within the same particle. To evaluate biological activity of bi-functional VLP a construction containing peptides representing dominant T- and B-cell epitopes of Asp f 2 was produced. Bi-functional particles were more potent in stimulating memory immune responses. These results demonstrate new possibilities of pl-380 based expression system to produce multifunctional VLPs.
State Research Institute for Genetics and Selection of Industrial Microorganisms, RAS, Moscow, Russia.
A N
AN
Marchenko
D G
DG
Kozlov
E V
EV
Svirshchevskaya
N Y
NY
Viskova
S V
SV
Benevolensky
D000595:Amino Acid Sequence; D000818:Animals; D001232:Aspergillus fumigatus; D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins; D007114:Immunization; D016056:Immunodominant Epitopes; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D018626:Retroelements / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014771:Virion / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*
D018985:Epitopes, B-Lymphocyte; D018984:Epitopes, T-Lymphocyte; D005656:Fungal Proteins; D016056:Immunodominant Epitopes; D008164:Luminescent Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins; D018626:Retroelements; C106810:allergen Asp f 2, Aspergillus fumigatus; C094737:p1 protein, yeast; D049452:Green Fluorescent Proteins