TBN1 is a nuclease with antitumor activity. The main goal of this work was to estimate how TBN1 and its modificated variants are stable in the ?leaf factory? system used for its production and whether it can be enhanced or influenced by chosen potential ?modificators? i. e. silencing supressors, transcription factors, glycosyltransferases and kinases. Nicotiana benthamina plants were infiltrated with the mixture of Agrobacterium tumefaciens strains bearing the nuclease plant expression vectors and co-infiltrated with the ?modifying? vectors. The nuclease and protein analyses revealed that nuclease TBN1 wt and its modificated variants are stable in the used ?leaf factory? system as to their molecular mass, only quantitative changes were detected. Expreximents showed that activity and production of the nucleases increased upon coinfiltration with silencing supressor and decreased upon coexpression with chosen transcription factor. Glycosyltransferases and kinases influenced activity and production only insignificantly. The experiments also revealed that modificated variants of TBN1 have different molecular weight suggesting that different N-glycosylation domains have different length of sugar chain and influence on nuclear activity. Our data show that this expression in planta seems to be suitable for production for study of antitumor activity of these nucleases.
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:48561 |
| Date | January 2011 |
| Creators | LOMNICKÁ, Anna |
| Source Sets | Czech ETDs |
| Language | Czech |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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