Expression of Human papillomavirus 16 E7ggg oncoprotein on N- and C-terminus of Potato virus X coat protein in bacterial and plant cells.
The E7 oncoprotein from Human papillomavirus type 16 (HPV16) is an attractive candidate for anti-cancer therapeutical vaccine development. In this study, we engineered different fusions of mutagenized coding sequence of E7 oncoprotein (E7ggg) with coat protein of Potato virus X (PVX CP) both on 5'- and 3'-terminus of PVX CP and evaluated the influence of the length of linker (no linker, 4, 15aa) connecting PVX CP and E7ggg on their production. At first the expression in Escherichia coli was conducted to assess the characteristics of the recombinant protein prior to be further produced in plants, that is, resultant proteins were used for screening of their immunological reactivity with antibodies against PVX CP and E7. Fusion proteins successfully expressed in bacteria and plants were partially purified and their reactivity and ability to form virus-like particles were evaluated with anti-E7 antibodies.
Academy of Sciences of the Czech Republic, 160 00 Prague 6, Czech Republic.
Helena
H
Plchova
Tomas
T
Moravec
Hana
H
Hoffmeisterova
Jitka
J
Folwarczna
Noemi
N
Cerovska
D024509:3' Flanking Region; D024506:5' Flanking Region; D016960:Agrobacterium tumefaciens; D000906:Antibodies / Q000276:immunology; D036022:Capsid Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D003001:Cloning, Molecular; D004926:Escherichia coli; D005260:Female; D015870:Gene Expression; D052162:Human papillomavirus 16 / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D050725:Papillomavirus E7 Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D030361:Papillomavirus Infections / Q000276:immunology / Q000821:virology; D053918:Papillomavirus Vaccines / Q000737:chemistry / Q000235:genetics*; D017863:Potexvirus / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D014026:Nicotiana; D002583:Uterine Cervical Neoplasms / Q000276:immunology / Q000821:virology; D058425:Vaccines, Virus-Like Particle / Q000737:chemistry / Q000235:genetics*
D000906:Antibodies; D036022:Capsid Proteins; D050725:Papillomavirus E7 Proteins; D053918:Papillomavirus Vaccines; D011993:Recombinant Fusion Proteins; D058425:Vaccines, Virus-Like Particle; C058750:coat protein, Potato virus X