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Figure 1 | Plant Methods

Figure 1

From: A procedure for localisation and electrophysiological characterisation of ion channels heterologously expressed in a plant context

Figure 1

Expression vectors engineered for transformation of tobacco mesophyll protoplasts. (A) pLoc. This vector carries a single over-expression cassette. The green fluorescent protein cDNA ("GFP") is framed by a strong promoter ("EN50PMA4") and a terminator ("T"). Three restriction sites (Bgl II, BamH I, Kpn I) are available for in-phase cloning of another cDNA. "ATG" indicates the first GFP codon. This vector allows the expression of fusion polypeptides with a GFP-tag in C-terminal for localisation purpose. (B) pFunct+Tag. This vector carries two expression cassettes both featuring the EN50PMA4 promoter and the T terminator. The first cassette encodes the GFP alone and the second one displays two restriction sites (Xho I, Sma I) for cDNA cloning. pFunct+Tag is used for functional expression of the recombinant cDNA while fluotagging transformed cells. (C) pFunct. This vector has the same construction as pFunct+Tag but the GFP cassette is absent. It is used, in combination of pLoc or pFunct+Tag, when two cDNAs are to be co-expressed (see text). EN50PMA4 is a tobacco enhanced promoter (see "Methods"); and T: is the nopaline synthase terminator of Agrobacterium tumefaciens. These vectors were obtained in a pTZ-19U plasmid (Stratagene, LaJolla, CA, USA) background.

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Plant Methods

ISSN: 1746-4811