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Fig. 3 TAP strategy and cloning vectors. A) The LAP-tagged target protein is first purified with anti-
GFP antibody coupled beads. LAP-tagged target protein and interacting proteins are released from the
beads by TEV protease cleavage and purified over S protein agarose. Interacting proteins are eluted from
the beads and analyzed by mass spectrometry. B) Schematic vector map illustrating the common features
of the standard LAP cloning vectors. C) Partial sequence of pIC26: in green 3 0 end of GFP, pink TEV
protease cleavage site, red S peptide. The SpeI cloning site is indicated. D) Overview of the different
features of the TAP cloning vectors. (See color plate.)
( Cheeseman et al., 2004 ). pIC26 contains pie-1 regulatory sequences to express the
LAP-tagged protein in the germline and embryo and can be integrated by ballistic
bombardment of the strain DP38 using unc-119 as a transformation marker (see
Section II.D.). pAA65 contains the same features as pIC26 but has mCherry instead
of GFP as a fluorescent tag ( Fig. 3B, D )( McNally et al., 2006 ). C. elegans LAP
vectors were reduced in size by introducing a truncated version of unc-119 in TH314
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