We applied the alkaline version of the single-cell gel electrophoresis (comet) assay to roots and leaves of tobacco (Nicotiana tabacum var. xanthi) seedlings or isolated leaf nuclei treated with: (1) the alkylating agent ethyl methanesulphonate, (2) necrotic heat treatments at 50 degrees C, and (3) DNase-I. All three treatments induced a dose-dependent increase in DNA migration, expressed as percentage of tail DNA. A comparison of the fluorochrome DNA dyes ethidium bromide, DAPI and YOYO-1 demonstrated that for the alkaline version of the comet assay in plants, the commonly used fluorescent dye ethidium bromide can be used with the same efficiency as DAPI or YOYO-1.
- MeSH
- alkylační látky toxicita MeSH
- analýza jednotlivých buněk MeSH
- barvení a značení metody MeSH
- benzoxazoly chemie MeSH
- chinolinové sloučeniny chemie MeSH
- deoxyribonukleasa I metabolismus farmakologie MeSH
- DNA rostlinná analýza chemie MeSH
- ethidium chemie MeSH
- ethylmethansulfonát toxicita MeSH
- fluorescenční barviva MeSH
- indoly chemie MeSH
- kometový test MeSH
- kořeny rostlin chemie účinky léků MeSH
- listy rostlin chemie účinky léků MeSH
- semenáček chemie účinky léků MeSH
- tabák chemie účinky léků MeSH
- vysoká teplota MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Vaccine strategies for the treatment of human papillomavirus-induced cervical cancer are based mainly on the human papillomavirus 16 E7 (HPV16 E7) oncoprotein. The immunogenicity of the E7 gene has been enhanced by its fusion to many different genes. Here, we linked a short sequence coding for the E7 peptide (aa 44-60) containing immunodominant epitopes for B and T cells to the 3' end of the gene coding for the whole coat protein (CP) of the poty-virus, potato virus A (PVA), and its deleted form (CPdel) with a short C-terminal deletion of 5 amino acids (LGVKG). CP-E7 and CPdel-E7 fusion proteins, just like CP alone, spontaneously assembled into virus-like particles in both procaryotic and eucaryotic cells. The CP-E7 and CPdel-E7 fusion genes induced slightly stronger E7-specific cytotoxic T-lymphocyte responses than the whole E7 gene, although they were still lower than those elicited by the previously constructed fusion gene, Sig/E7GGG/LAMP-1. The E7- and CP-specific antibody responses were not detected in mice vaccinated with CP-E7 and CPdel-E7 fusion genes. The CP-E7 and CPdel-E7 fusion genes protected mice against the development of tumors induced by TC-1 cells producing the E7 antigen and were also effective in the therapeutic setting, i.e. when the vaccination was performed after tumor cell administration. Their antitumor effect was comparable to those of the whole E7 gene and Sig/E7GGG/LAMP-1 fusion gene. There was no relevant difference between immune responses elicited by CP-E7 and CPdel-E7 DNA vaccination.
- MeSH
- antitumorózní látky farmakologie MeSH
- biolistika MeSH
- buněčné linie MeSH
- buňky NIH 3T3 MeSH
- časové faktory MeSH
- cytotoxické T-lymfocyty cytologie MeSH
- DNA vakcíny MeSH
- ELISA MeSH
- financování organizované MeSH
- genetická terapie metody MeSH
- hmotnostní spektrometrie MeSH
- lidé MeSH
- molekulární sekvence - údaje MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- nádorové buněčné linie MeSH
- onkogenní proteiny virové chemie MeSH
- peptidy chemie MeSH
- plazmidy metabolismus MeSH
- Potyvirus genetika MeSH
- protinádorové vakcíny MeSH
- rekombinantní fúzní proteiny chemie MeSH
- RNA chemie MeSH
- sekvence aminokyselin MeSH
- sekvenční homologie aminokyselin MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- technika přenosu genů MeSH
- transmisní elektronová mikroskopie MeSH
- transplantace nádorů MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- MeSH
- diabetické angiopatie diagnóza MeSH
- dolní končetina MeSH
- lidé MeSH
- pletyzmografie impedanční MeSH
- Check Tag
- lidé MeSH
