Abstract
E2Tag has been used for tagging of bacterial, yeast, and mammalian proteins. A set of plant expression vectors was constructed for tagging proteins with E2Tag. Detection of E2-tagged proteins with specific antibodies inNicotiana benthamiana andNicotiana tabacum was easily done without any nonspecific background activity. No effect on the biological activity of the enzyme GUS, used as a marker, was detected. The subcellular localization of E2-tagged plant protein AtRLI2 with yet unknown functions was described.
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Abbreviations
- 2–5A:
-
2′,5′-oligoadenylate
- 35S:
-
cauliflower mosaic virus 35S promoter
- E2Tag:
-
novel epitope derived from a bovine papillomavirus type 1 transactivator protein E2
- GUS:
-
β-glucuronidase
- MES:
-
2(N-morpholino)ethanesulfonic acid
- nos :
-
nopaline synthase gene from theAgrobacterium tumefaciens Ti plasmid
- PBS:
-
phosphate-buffered saline
- PCR:
-
polymerase chain reaction
- RLI:
-
RNase L inhibitor
- uidA :
-
Escherichia coli gene encoding β-glucuronidase
- X-Gluc:
-
5-bromo-4-chloro-3-indolyl-beta-D-glucuronic acid
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Nigul, L., Olspert, A., Meier, M. et al. New plant vectors for protein tagging with E2 epitope. Plant Mol Biol Rep 22, 399–407 (2004). https://doi.org/10.1007/BF02772682
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DOI: https://doi.org/10.1007/BF02772682